This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fernández, S.
Right arrow Articles by Alonso, J. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fernández, S.
Right arrow Articles by Alonso, J. C.

 Previous Article  |  Next Article 

Vol. 180, Issue 13, 3405-3409, July 1, 1998

Genetic Recombination in Bacillus subtilis 168: Effects of recU and recS Mutations on DNA Repair and Homologous Recombination

Silvia Fernández1, Alexei Sorokin2, and Juan C. Alonso1*

1 Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas, Campus Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain,1 and 2 Institut National de la Recherche Agronomique---Génétique Microbienne, 78352 Jouy-en-Josas Cedex, France2

    ABSTRACT
Top
Abstract
Introduction
Materials & Methods
Results & Discussion
References

Bacillus subtilis recombination-deficient mutants were constructed by inserting a selectable marker (cat gene) into the yppB and ypbC coding regions. The yppB:cat and ypbC:cat null alleles rendered cells sensitive to DNA-damaging agents, impaired plasmid transformation (25- and 100-fold), and moderately affected chromosomal transformation when present in an otherwise Rec+ B. subtilis strain. The yppB gene complemented the defect of the recG40 strain. yppB and ypbC and their respective null alleles were termed "recU" and "recU1" (recU:cat) and "recS" and "recS1" (recS:cat), respectively. The recU and recS mutations were introduced into rec-deficient strains representative of the alpha  (recF), beta  (addA5 addB72), gamma  (recH342), and varepsilon  (recG40) epistatic groups. The recU mutation did not modify the sensitivity of recH cells to DNA-damaging agents, but it did affect inter- and intramolecular recombination in recH cells. The recS mutation did not modify the sensitivity of addAB cells to DNA-damaging agents, and it marginally affected recF, recH, and recU cells. The recS mutation markedly reduced (about 250-fold) intermolecular recombination in recH cells, and there were reductions of 10- to 20-fold in recF, addAB, and recU cells. Intramolecular recombination was blocked in recS recF, recS addAB, and recS recU cells. RecU and RecS have no functional counterparts in Escherichia coli. Altogether, these data indicate that the recU and recS proteins are required for DNA repair and intramolecular recombination and that the recF (alpha  epistatic group), addAB (beta ), recH (gamma ), recU (varepsilon ), and recS genes provide overlapping activities that compensate for the effects of single mutation. We tentatively placed recS within a new group, termed "zeta ."

    INTRODUCTION
Top
Abstract
Introduction
Materials & Methods
Results & Discussion
References

In Bacillus subtilis, several recombination-deficient mutants with increased sensitivities to DNA-damaging agents have been isolated. Genetic analysis of chromosomal and plasmid transformations has demonstrated that to be efficient, homologous recombination requires the products of the recA, recB, recD, recF, recU (formerly termed "recG"), recH, recL, recN, recP, recR, addA, addB, and mfd genes (reviewed in reference 6), in addition to the enzymes involved in general DNA metabolism, such as DNA polymerases, SSB, Hbsu, DNA ligase, and topoisomerases, among others (15, 20).

Three qualitatively distinct situations may arise when the frequency of genetic exchange of a double-mutant strain is analyzed. (i) The frequency may be equal to that of the more deficient single-mutant parent. (ii) The frequency may be equal to the sum of each of the single-mutant parents (additive effect). (iii) The frequency may be greater than the sum of each of the single-mutant parents (synergistic effect) (11). Considering these possibilities, we have classified the B. subtilis double-Rec- strains into different epistatic groups (alpha , beta , gamma , and varepsilon  groups) (3). The alpha  epistatic group activity requires the recF, recL, recR, and recN genes (2). Recently, the RecF and RecR proteins have been biochemically characterized (7-9). The beta  epistatic group activity is dependent on the addA and addB genes (3). The addA and addB genes encode different subunits of the multifunctional enzyme AddAB (also termed exonuclease V [Exo V] or RecBCD in Escherichia coli [6, 19]). The gamma  epistatic group activity requires the recP and recH genes (6). The varepsilon  epistatic group activity may require the recB, recD, and recU genes (6). The gamma  and varepsilon  epistatic groups, which are poorly characterized, do not seem to have a counterpart in E. coli. No double-mutant strains impaired in the functions classified within groups gamma  and varepsilon  have been described.

In this report, we genetically characterize the products of the B. subtilis recU (which codes for an unknown activity) and recS (which codes for a putative DNA helicase) genes that are involved in DNA repair and homologous recombination. The phenotype associated with recU cells is consistent with our previous grouping, but we were unable to classify recS cells within any previously recognized epistatic group (see reference 6). Unless otherwise stated, the indicated genes and products are of B. subtilis origin.

    MATERIALS AND METHODS
Top
Abstract
Introduction
Materials & Methods
Results & Discussion
References

Bacterial strains and plasmids. The E. coli JM103 strain was used (23). All B. subtilis strains listed in Table 1 are isogenic with strain YB886 (rec+ control). The ypbC (National Biomedical Research Foundation-Protein Identification Resources [NBRF-PIR] and Swiss-Prot accession no. .Gb_Ba) and yppB (.Gb_Ba) genes were PCR amplified with specific oligonucleotides and cloned (Table 1). The transfer of the recU:cat deletion/insertion allele (termed recU1) and recS:cat deletion/insertion allele (termed recS1) from pCB182 and pCB176, respectively, into the B. subtilis chromosome by a double-crossover event was performed as previously described (5).

                              
View this table:
[in this window]
[in a new window]
 
Table 1
Plasmids and bacterial strains used in this study

Chemical treatment. Methyl methanesulfonate (MMS) was purchased from Eastman Kodak, Rochester, N.Y. 4-Nitroquinoline-1-oxide (4NQO) was purchased from Sigma Chemical Co., St. Louis, Mo. The chemical treatment of the mutant strains was performed essentially as previously described (1).

Transformation of bacteria. Competent cells were prepared as described by Rottländer and Trautner (25) and Maniatis et al. (23) for B. subtilis and E. coli, respectively. Plasmid transformants were selected on tryptone-yeast agar medium (25) containing chloramphenicol or erythromycin at 5 µg/ml. Met+ recombinants were selected by plating on minimal agar containing all nutritional requirements except methionine (25).

Recombination frequencies. Relative transformation frequencies were used as a measure of recombination. B. subtilis competent cells (about 5.0 × 107 cells/ml) were transformed according to the method of Rottländer and Trautner (25). The yield of Met+ or plasmid transformants was corrected for DNA uptake and cell viability as described by Alonso et al. (1). DNA uptake, which is taken as a measurement of competence, was monitored as described previously (25).

    RESULTS AND DISCUSSION
Top
Abstract
Introduction
Materials & Methods
Results & Discussion
References

Replacement of B. subtilis recU and recS genes by use of a selectable marker. Recently by molecular cloning, DNA sequencing, and computer homology comparisons, a B. subtilis open reading frame (ORF), ypbC, was identified as coding for a putative recombinational helicase, the gene for which is "recQ-like" (coordinates 4291 to 5781 in the sequence with accession no. .Gb_Ba) (27). Furthermore, a B. subtilis region (plasmid pMP29-42) able to rescue the recG40 mutation (24) has been sequenced. The pMP29-42-borne DNA segment bears two truncated ORFs, termed "yppB" and "yppC" (27) (accession no. .Gb_Ba).

To identify and clone the recG gene of B. subtilis, a PCR-amplified DNA with coding capacity for yppB or yppC was used to transform the recG40 strain. Upon transformation of the MMS-sensitive (MMSs) recG40 strain with purified DNA fragments, we learned that only the yppB ORF fully complements the recG40 defect, since yppB encodes a protein unrelated to the E. coli recG product. To avoid confusion in the nomenclature and for the sake of simplicity, we have renamed this ORF as "recU" and the mutant allele as "recU40."

To learn whether the putative ypbC gene codes for a product involved in homologous recombination and recombinational repair and to challenge the grouping of the B. subtilis rec genes (described above), we cloned the chloramphenicol acetyltransferase gene (cat) 239 and 415 bp downstream of the ATG codons of the ypbC and recU genes, respectively. The resulting alleles were used to replace the ypbC and recU genes.

Strains containing the expected gene substitution were obtained by natural transformation of the B. subtilis rec+ strain and its isogenic rec-deficient derivatives with linearized plasmid-borne recU:cat (recU1) and ypbC:cat (Table 1). The presence of the desired replacement was confirmed by PCR amplification and nucleotide sequence analysis (data not shown). In all cases, integration had occurred by a double-crossover event, as predicted for transformation of competent cells with linear plasmid DNA (reviewed in reference 13).

To analyze if the constructs were impaired in recombinational repair processes, we exposed the mutant strains to the lethal effect of MMS. Both recU1 and ypbC:cat strains were unable to form colonies on plates containing 300 (2.7 mM) and 100 (0.9 mM) µg of MMS per ml, respectively, whereas the wild-type strain formed colonies even in plates containing 450 µg (4 mM) of MMS per ml. The recU1 and ypbC:cat strains, exposed to the killing action of 10 mM MMS, were more sensitive than the otherwise rec+ strain (Fig. 1A and Fig. 2). The same results were obtained when the cells were challenged with 0.1 mM 4NQO (data not shown). It is likely, therefore, that the ypbC gene product is involved in DNA repair. We have renamed the recS gene and its mutant allele "recS1" (ypbC:cat). As revealed in Fig. 1A, the cells carrying the recU1 null allele are slightly more sensitive to the killing action of MMS than those of the recU40 strain, and the defect can be complemented by a plasmid-borne recU gene (coordinates 2536 to 2874 in the sequence with accession no. .Gb_Ba).


View larger version (12K):
[in this window]
[in a new window]
 
Fig. 1.   Survival of B. subtilis strains following exposure to 10 mM MMS. (A) Survival of the rec+ strain (solid circles), the recU40 strain (open circles), the recU40 strain carrying a plasmid-borne recU gene (solid triangles), and the recU1 strain (solid squares). (B) Survival of rec+ (solid circles), recH342 (open squares), recU1 (solid squares), and recH342 recU1 (solid triangles) strains. The chemical treatment of DNA repair-deficient mutant strains was performed essentially as previously described (1).

Effect of DNA-damaging agents in recU and recH mutant strains. To date in B. subtilis, 13 genes involved in homologous recombination (recA, addA, addB, recF, recL, recR, recN, recH, recP, recB, recD, recU, and mfd) have been identified (reviewed in reference 6). Genetic analysis of double-rec mutants with chromosomal and plasmid transformation has led to the classification of the rec genes, other than recA, into four epistatic groups (3). The genes involved in the alpha  (recF, recL, recR, and recN) and beta (addA and addB) groups are well characterized (7, 8, 19), but those comprising the gamma  (recH and recP) and varepsilon  (recB, recD, and recU) groups are poorly defined.

Previously, we have shown that the recU40 (varepsilon  group) gene in combination with mutations in the addA addB (beta  epistatic group) genes was more sensitive to the killing action of MMS and 4NQO than the single-mutant parent, whereas with a mutation in the recF (alpha  group) gene, it was as sensitive as the single parent (3, 5). However, no data are available about the effect of double mutations in the gamma  and varepsilon  groups. The recH342 strain was selected as a representative of the gamma  epistatic group. We have constructed a recU recH double mutant and challenged these cells with the killing action of 10 mM MMS. As revealed in Fig. 1B, the recH mutation did not affect the MMS sensitivity of the recU cells.

Effects of recS, recF, recH, recU, and addAB mutations on MMS sensitivity. B. subtilis rec+ and its isogenic rec-deficient derivative strains (BG431, BG435, BG429, and BG433 [listed in Table 1]) were exposed to the killing action of 10 mM MMS. The recF, addAB, recH, and recU genes were selected as representatives of the alpha , beta  gamma , and varepsilon  groups, respectively. As revealed in Fig. 2, various degrees of increased sensitivity were observed. The recS, addAB, and recH strains are moderately affected, whereas the recF and recU strains are very sensitive to the killing action of MMS compared to the rec+ control (Fig. 2). The recS mutation partially suppressed the sensitivity of addAB cells to MMS (Fig. 2A). The recS mutation moderately increased the MMS sensitivity of recH, recU, and recF cells (Fig. 2B, C, and D).


View larger version (17K):
[in this window]
[in a new window]
 
Fig. 2.   Survival of B. subtilis strains following exposure to 10 mM MMS. (A) Survival of rec+ (solid circles), recS1 (open circles), addA addB (solid squares), and addA addB recS1 (solid triangles) strains. (B) Survival of rec+ (solid circles), recS1 (open circles), recH342 (solid squares), and recH342 recS1 (solid triangles) strains. (C) Survival of rec+ (solid circles), recS1 (open circles), recF15 (solid squares), and recF15 recS1 (solid triangles) strains. (D) Survival of rec+ (solid circles), recS1 (open circles), recU40 (solid squares), and recU40 recS1 (solid triangles) strains.

Together, these results indicate that (i) the recS product is needed for the removal of DNA damage; (ii) the recS genes contribute to different extents to the removal of DNA damage in recH, recU, and recF cells; and (iii) recS partially suppresses the defect of the addAB cells.

Effects of recF, addAB, recU, recH, and recS mutations on genetic recombination. To study the effects of the recF, addAB, recH, recU, and recS mutations on genetic recombination, we analyzed the requirement for these functions in natural transformational recombination. We measured the frequency of transformation by using plasmid and chromosomal DNA. Transformation in B. subtilis involves the transfer of naked double-stranded DNA from the media to the recipient competent uninucleated cell (22, 26). During DNA uptake, a set of competence (com) gene products degrades one of the DNA strands and takes up the other one in a linear single-stranded form (reviewed in reference 13). Therefore, by the activity of the com genes, the donor DNA is presented to the recombinational machinery in a form such that it is ready for homology searching (synaptic stage) on the parental molecule (13, 22). Hence, in our analysis, we are studying the synaptic and postsynaptic stages but neglect the involvement of rec functions in the presentation of the substrate (presynaptic stage).

B. subtilis transformation with chromosomal DNA (chromosomal transformation) does not require replication for integration of donor markers (13, 22). In contrast, both replication and recombination functions are required for the establishment of plasmid DNA (13, 22). Canosi et al. (12) and de Vos et al. (12a) have proposed that after uptake of the oligomeric plasmid DNA molecule (a monomer is inactive in transformation) and synthesis of the complementary strand, pairing of one of the incoming single-stranded DNA ends with the newly replicated strand results in plasmid establishment. By measuring both chromosomal (intermolecular recombination) and plasmid (intramolecular recombination) transformation, we can examine different types of events. B. subtilis competent cells were transformed with 1 µg of homologous chromosomal DNA or plasmid DNA per ml to determine the transformation frequency of the Rec- mutant strains. The frequency of appearance of met+ transformants in the single-Rec- strain and in certain double-Rec- strains has been previously reported (1, 3). Here, the experiments were performed in parallel for comparison with other strains (Table 2).

                              
View this table:
[in this window]
[in a new window]
 
Table 2
Effect of RecU and RecS on homologous recombination as measured by transformation of chromosomal and plasmid DNAa

Except for recA, the frequency of chromosomal transformation (intermolecular recombination) in the singly rec-deficient strain did not change more than fourfold relative to the rec+ value (3) (Table 2). The same results were found for plasmid transformation.

The recU null allele (recU1) and recU40 mutations reduce the chromosomal transformation less than threefold (1) (Table 2). As previously reported, plasmid transformation is marginally affected by the recU40 mutation, but it is reduced 25-fold and 100-fold in the recU1 and recS1 cells, respectively (Table 2).

Since (i) the recH (epistatic group gamma ) mutation blocked (>5,000-fold reduction) chromosomal transformation of recF (group alpha ) and addAB (group beta ) cells (3, 5) and reduced about 250-fold that of recS cells (Table 2) but did not affect chromosomal transformation of recU cells (group varepsilon ), and (ii) the recH mutation blocks plasmid transformation of recF addAB cells (5) and reduced more than 1,000-fold that of recS cells (Table 2) but did not affect (2-fold) plasmid transformation of recU cells (Table 2), we can classify recH and recS into two different epistatic groups by using transformational recombination, and confirmed our previous classification of recH and recU within different epistatic groups (3). The classification of recH within group gamma  and recU within varepsilon  was based on the following facts. (i) recH cells are only moderately sensitive to DNA-damaging agents, whereas recU cells are very sensitive to these agents (1, 3). (ii) Double-mutant strains impaired in the alpha  plus gamma  or beta  plus gamma  epistatic groups are blocked in chromosomal recombination, whereas the alpha  plus varepsilon  or beta  plus varepsilon  groups are moderately affected (1, 3). (iii) Transductional intramolecular recombination is modestly affected in the double alpha  plus gamma  mutants but is highly impaired in the alpha  plus varepsilon  mutants (4). (iv) Chromosomal transformation is blocked in hbs recH cells but is reduced only 11-fold in hbs recU40 cells (15). To reconcile these differences with the data presented in this report, we have to assume that certain gene products of the gamma  and varepsilon  epistatic groups cooperate and that recH and recU operate at different stages in the recombination pathway. Both the gamma  and varepsilon  groupings will tentatively be maintained until more information becomes available.

The frequency of chromosomal transformation of the recS null allele was reduced about 4-fold, whereas plasmid transformation was reduced about 100-fold (Table 2). The recS mutation reduced chromosomal transformation in recF, recU, and addAB cells about 10- to 20-fold and that in recH cells about 250-fold (Table 2). Plasmid transformation was reduced by the recS mutation more than 1,000-fold in strains representative of the beta , gamma , and varepsilon  epistatic groups and was reduced about 200-fold in the alpha  epistatic group.

On the basis of these and previous results, we conclude that no interaction exists between the RecU protein and the addAB and recF gene products, whereas a significant interaction may exist between the RecU and RecH proteins. No interaction was observed between the recS and the addAB, recH, recU, and recF functions. We tentatively place the recS mutation within a different epistatic group, termed "zeta ."

RecU and RecS proteins have no functional counterparts in E. coli. A homology search of the RecU and RecS proteins was performed with the protein sequences available in the NBRF-PIR (release 50) and Swiss-Prot (release 34) databases. Significant identity (about 50%) was observed between the 23.9-kDa RecU protein (database accession no. ) and an equivalent 23.1-kDa product of Streptococcus pneumoniae (27) (accession no. ypoa_strpn, ypoa_stror), and 34% identity was observed with a 19.3-kDa protein of Mycoplasma genitalium (17) (accession no. ). A protein similar to RecU has not been detected so far in gram-negative bacteria. Both E. coli and Haemophilus influenzae seem to lack this protein (10, 16).

The 56.5-kDa RecS protein (accession no. .GB_Ba) shares 34 to 36% identity with E. coli RecQ (27) (accession no. BVECRQ) and the putative H. influenzae RecQ (accession no. U32756_D) and B. subtilis "RecQ" (accession no. AF027868.Gb_Ba) proteins. The degree of identity of RecS with both E. coli RecQ and the putative B. subtilis "RecQ" protein could be enhanced up to 43 and 40%, respectively, if only the regions containing the seven-amino-acid motifs of DExH-box DNA helicases (first 330 residues) were used for the alignment. The putative B. subtilis "RecQ" protein (yocI) shares 40% identity with the E. coli RecQ protein.

Unlike E. coli containing only one recQ gene product with DNA helicase activity (28), B. subtilis could possess more than one such putative helicase (21). This is consistent with the fact that human cells contain genes that code for three related putative helicases (RecQL [human RecQL], accession no. A55311), HumBS [Bloom's syndrome, accession no. A57570], and HumWS [Werner's syndrome, accession no. L76937]) (see reference 14). Although the B. subtilis "recQ" gene has not yet been tested, the gene seems to be nonessential for cell viability (reviewed in references 14 and 20).

It is generally accepted that the role of E. coli RecQ (and probably B. subtilis "RecQ") protein, alone or in combination with the E. coli RecJ protein, is to generate a 3'-terminal single-stranded DNA to which RecA could bind in a step proceeding to the homologous pairing. With chromosomal and plasmid transformation, we are measuring the synaptic and postsynaptic stages of recombination (13, 22). In this report, we show that the RecS protein is required in either the synaptic stage, postsynaptic stage, or both stages. It remains to be documented if RecS is a bona fide DNA helicase and if the helicase activity participates directly in recombination. We also report here that deletion of the recS gene partially suppressed the defect of addAB cells. The AddAB protein is a multifunctional enzyme associated with nuclease and helicase activities (19, 20); however, the defects generated by the addA5 and addB71 mutations in this genetic background remain to be characterized.

    ACKNOWLEDGMENTS

This research was partially supported by grant PB 96-0817 from DGCICYT and grant 06G/004/96 from the Consejería de Educación y Cultura de la Comunidad de Madrid to J.C.A. and grants from GREG (décision 21) and ECC (BIO2-CT93-0272 and BIO2-CT94-2011) to S. D. Ehrlich. S.F. is supported by a Comunidad de Madrid training grant.

    FOOTNOTES

* Corresponding author. Mailing address: Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone: (34) 91 585 4546. Fax: (34) 91 585 4506. E-mail: jcalonso{at}cnb.uam.es.

    REFERENCES
Top
Abstract
Introduction
Materials & Methods
Results & Discussion
References

  1. Alonso, J. C., Tailor, R. H., and Lüder, G. (1988) Characterization of recombination-deficient mutants of Bacillus subtilis. J. Bacteriol. 170, 3001-3007[Abstract/Free Full Text].
  2. Alonso, J. C., and Lüder, G. (1991) Characterization of recF suppressors in Bacillus subtilis. Biochimie 73, 277-280[Medline].
  3. Alonso, J. C., Lüder, G., and Tailor, R. H. (1991) Characterization of the Bacillus subtilis recombinational pathways. J. Bacteriol. 173, 3977-3980[Abstract/Free Full Text].
  4. Alonso, J. C., Lüder, G., and Trautner, T. A. (1992) Intramolecular homologous recombination in Bacillus subtilis 168. Mol. Gen. Genet. 236, 60-64[Medline].
  5. Alonso, J. C., Stiege, A. C., and Lüder, G. (1993) Molecular analysis of the Bacillus subtilis recF function. Mol. Gen. Genet. 239, 129-136[Medline].
  6. Alonso, J. C., Ayora, S., and Rojo, F. (1996) Recombinación genética en Bacillus subtilis in Microbiología y genética molecular. (Casadesús, J., ed), pp. 229-240, Publicaciones Universidad de Huelva, Huelva, Spain.
  7. Ayora, S., and Alonso, J. C. (1997) Purification and characterization of the RecF protein from Bacillus subtilis 168. Nucleic Acids Res. 25, 2766-2772[Abstract/Free Full Text].
  8. Ayora, S., Stiege, A. C., and Alonso, J. C. (1997) RecR is a zinc metalloprotein from Bacillus subtilis 168. Mol. Microbiol. 23, 639-647[Medline].
  9. Ayora, S., Stiege, A. C., Lurz, R., and Alonso, J. C. (1997) Bacillus subtilis 168 RecR protein-DNA complexes visualized as looped structures. Mol. Gen. Genet. 254, 54-62[Medline].
  10. Blattner, F. R., Plunkett, G., Bloch, C. A., Perna, N. T., Burland, V., Riley, M., Collado-Vides, J., Glasner, J. D., Rode, C. H., Mayhew, G. F., Gregor, J., Davis, N. W., Kirkepatrick, H. A., Goeden, M. A., Rose, D. J., Man, B., and Shao, Y. (1997) The complete genome sequence of Escherichia coli K-12. Science 277, 1453-1462[Abstract/Free Full Text].
  11. Brendel, M., and Haynes, R. H. (1973) Interactions among genes controlling sensitivity to radiation and alkylation in yeast. Mol. Gen. Genet. 125, 197-216[Medline].
  12. Canosi, U., Iglesias, A., and Trautner, T. A. (1981) Plasmid transformation in Bacillus subtilis: effects of insertion of Bacillus subtilis DNA into plasmid pC194. Mol. Gen. Genet. 181, 434-440[Medline].
  13. de Vos, W. M., Venema, G., Canosi, U., and Trautner, T. A. (1981) Plasmid transformation in Bacillus subtilis: fate of plasmid DNA. Mol. Gen. Genet. 181, 424-433[Medline].
  14. Dubnau, D. (1993) Genetic exchange and homologous recombination in Bacillus subtilis and other gram-positive bacteria: biochemistry, physiology, and molecular genetics. (Sonenshein, A. L., Hoch, J. A., and Losick, R., eds), pp. 555-584, American Society for Microbiology, Washington, D.C.
  15. Ellis, N. A., and German, J. (1996) Molecular genetics of Bloom's syndrome. Hum. Mol. Genet. 5, 1457-1463[Abstract].
  16. Fernández, S., Rojo, F., and Alonso, J. C. (1997) The Bacillus subtilis chromatin-associated protein Hbsu is involved in DNA repair and recombination. Mol. Microbiol. 23, 1169-1179[Medline].
  17. Fleischmann, R. D., Adams, M. D., White, O., Clayton, R. A., Kirkness, E. F., Kerlavage, A. R., Bult, C. J., Tomb, J. F., Dougherty, B. A., Merrick, J. M., McKenney, K., Sutton, G., FitzHugh, W., Fields, C., Gocayne, J. D., Scott, J., Shirley, R., Liu, L., Glodek, A., Kelley, J. M., Weidman, J. F., Phillips, C. A., Springs, T., Hedblom, E., Cotton, M. D., Utterback, T. R., Hanna, M. C., Nguyen, D. T., Saudek, D. M., Bramdon, R. C., Fine, L. D., Fritchman, J. L., Fuhrmann, J. L., Geoghagen, N. S. M., Gnehm, C. L., McDonald, L. A., Small, K. V., Fraiser, C. M., Smith, H. O., and Venter, J. C. (1995) Whole genome random sequencing and assembly of Haemophilus influenzae Rd. Science 269, 496-512[Abstract/Free Full Text].
  18. Fraser, C. D., Gocayne, J. D., White, O., Adams, M. D., Clayton, R. A., Fleischmann, R. D., Bult, C. J., Kerlavage, A. R., Sutton, G., Kelley, J. M., Fritchman, J. L., Weidman, J. F., Small, K. V., Sandusky, M., Fuhrmann, J., Nguyen, D., Utterback, T. R., Saudek, D. M., Phillips, C. A., Merrick, J. M., Tomb, J. F., Dougherty, B. A., Bott, K. F., Hu, P., Lucier, T. S., Peterson, S. N., Smith, H. O., Hutchinson III, C. A., and Venter, J. C. (1995) The minimal gene complement of Mycoplasma genitalium. Science 270, 397-403[Abstract/Free Full Text].
  19. Haima, P., Bron, S., and Venema, G. (1987) The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg. Mol. Gen. Genet. 209, 335-342[Medline].
  20. Kooistra, J., and Venema, G. (1991) Cloning, sequencing, and expression of Bacillus subtilis genes involved in ATP-dependent nuclease synthesis. J. Bacteriol. 173, 3644-3655[Abstract/Free Full Text].
  21. Kowalczykowski, S. C., Dixon, D. A., Eggleston, A. K., Lauder, S. D., and Rehrauer, W. M. (1994) Biochemistry of homologous recombination in Escherichia coli. Microbiol. Rev. 58, 401-465[Abstract/Free Full Text].
  22. Kunst, F., Ogasawara, N., Moszer, I., Albertini, A. M., Alloni, G., Azevedo, V., Bestero, M. G., et al.. (1997) The complete genome sequence of the Gram-positive model organism Bacillus subtilis. Nature 390, 249-256[Medline].
  23. Lacks, S. A. (1988) Mechanisms of genetic recombination in gram-positive bacteria in Genetic recombination. (Kucherlapati, R., and Smith, G. R., eds), pp. 43-86, American Society for Microbiology, Washington, D.C.
  24. Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982) Molecular cloning: a laboratory manual., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
  25. Perego, M., Ferrari, E., Bassi, M. T., Galizzi, A., and Mazza, P. (1987) Molecular cloning of Bacillus subtilis genes involved in DNA metabolism. Mol. Gen. Genet. 209, 8-14[Medline].
  26. Rottländer, E., and Trautner, T. A. (1970) Genetic and transfection studies with B. subtilis phage SP50. I. Phage mutants with restricted growth on B. subtilis strain 168. Mol. Gen. Genet. 108, 47-60[Medline].
  27. Singh, R. N., and Pitale, M. P. (1967) Enrichment of Bacillus subtilis transformants by zonal centrifugation. Nature 213, 1262-1263.
  28. Sorokin, A., Azevedo, V., Zumstein, E., Galleron, N., Ehrlich, S. D., and Serror, P. (1996) Sequence analysis of the Bacillus subtilis chromosome region between the serA and kdg loci cloned in a yeast artificial chromosome. Microbiology 142, 2005-2016[Abstract/Free Full Text].
  29. Umezu, K., and Kolodner, R. D. (1994) Protein interactions in genetic recombination in Escherichia coli. Interactions involving RecO and RecR overcome the inhibition of RecA by single-stranded DNA-binding protein. J. Biol. Chem. 269, 30005-30013[Abstract/Free Full Text].
  30. Yasbin, R. E., Field, P. I., and Anderson, B. J. (1980) Properties of Bacillus subtilis 168 derivates freed of their natural prophages. Gene 12, 155-159[Medline].


Copyright © 1998 by American Society for Microbiology


This article has been cited by other articles:

  • Cardenas, P. P., Carrasco, B., Sanchez, H., Deikus, G., Bechhofer, D. H, Alonso, J. C (2009). Bacillus subtilis polynucleotide phosphorylase 3'-to-5' DNase activity is involved in DNA repair. Nucleic Acids Res 37: 4157-4169 [Abstract] [Full Text]  
  • Dillingham, M. S., Kowalczykowski, S. C. (2008). RecBCD Enzyme and the Repair of Double-Stranded DNA Breaks. Microbiol. Mol. Biol. Rev. 72: 642-671 [Abstract] [Full Text]  
  • Canas, C., Carrasco, B., Ayora, S., Alonso, J. C. (2008). The RecU Holliday junction resolvase acts at early stages of homologous recombination. Nucleic Acids Res 36: 5242-5249 [Abstract] [Full Text]  
  • Sharma, S., Stumpo, D. J., Balajee, A. S., Bock, C. B., Lansdorp, P. M., Brosh, R. M. Jr., Blackshear, P. J. (2007). RECQL, a Member of the RecQ Family of DNA Helicases, Suppresses Chromosomal Instability. Mol. Cell. Biol. 27: 1784-1794 [Abstract] [Full Text]  
  • Sanchez, H., Kidane, D., Castillo Cozar, M., Graumann, P. L., Alonso, J. C. (2006). Recruitment of Bacillus subtilis RecN to DNA Double-Strand Breaks in the Absence of DNA End Processing. J. Bacteriol. 188: 353-360 [Abstract] [Full Text]  
  • Lopez-Torrejon, G., Martinez-Jimenez, M. I., Ayora, S. (2006). Role of LrpC from Bacillus subtilis in DNA transactions during DNA repair and recombination. Nucleic Acids Res 34: 120-129 [Abstract] [Full Text]  
  • Sanchez, H., Kidane, D., Reed, P., Curtis, F. A., Cozar, M. C., Graumann, P. L., Sharples, G. J., Alonso, J. C. (2005). The RuvAB Branch Migration Translocase and RecU Holliday Junction Resolvase Are Required for Double-Stranded DNA Break Repair in Bacillus subtilis. Genetics 171: 873-883 [Abstract] [Full Text]  
  • Carrasco, B., Cozar, M. C., Lurz, R., Alonso, J. C., Ayora, S. (2004). Genetic Recombination in Bacillus subtilis 168: Contribution of Holliday Junction Processing Functions in Chromosome Segregation. J. Bacteriol. 186: 5557-5566 [Abstract] [Full Text]  
  • Duez, C., Hallut, S., Rhazi, N., Hubert, S., Amoroso, A., Bouillenne, F., Piette, A., Coyette, J. (2004). The ponA Gene of Enterococcus faecalis JH2-2 Codes for a Low-Affinity Class A Penicillin-Binding Protein. J. Bacteriol. 186: 4412-4416 [Abstract] [Full Text]  
  • Ayora, S., Carrasco, B., Doncel, E., Lurz, R., Alonso, J. C. (2004). Bacillus subtilis RecU protein cleaves Holliday junctions and anneals single-stranded DNA. Proc. Natl. Acad. Sci. USA 101: 452-457 [Abstract] [Full Text]  
  • Carrasco, B., Fernandez, S., Petit, M.-A., Alonso, J. C. (2001). Genetic Recombination in Bacillus subtilis 168: Effect of {Delta}helD on DNA Repair and Homologous Recombination. J. Bacteriol. 183: 5772-5777 [Abstract] [Full Text]  
  • Autret, N., Dubail, I., Trieu-Cuot, P., Berche, P., Charbit, A. (2001). Identification of New Genes Involved in the Virulence of Listeria monocytogenes by Signature-Tagged Transposon Mutagenesis. Infect. Immun. 69: 2054-2065 [Abstract] [Full Text]  
  • Sciochetti, S. A., Piggot, P. J., Blakely, G. W. (2001). Identification and Characterization of the dif Site from Bacillus subtilis. J. Bacteriol. 183: 1058-1068 [Abstract] [Full Text]  
  • Pedersen, L. B., Setlow, P. (2000). Penicillin-Binding Protein-Related Factor A Is Required for Proper Chromosome Segregation in Bacillus subtilis. J. Bacteriol. 182: 1650-1658 [Abstract] [Full Text]  
  • Bolotin, A., Wincker, P., Mauger, S., Jaillon, O., Malarme, K., Weissenbach, J., Ehrlich, S. D., Sorokin, A. (2001). The Complete Genome Sequence of the Lactic Acid Bacterium Lactococcus lactis ssp. lactis IL1403. Genome Res 11: 731-753 [Abstract] [Full Text]  

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Fernández, S.
Right arrow Articles by Alonso, J. C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fernández, S.
Right arrow Articles by Alonso, J. C.