This Article
Right arrow Full Text
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 De Las Rivas, B.
Right arrow Articles by García, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by De Las Rivas, B.
Right arrow Articles by García, P.

 Previous Article  |  Next Article 

Journal of Bacteriology, September 2002, p. 4988-5000, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.4988-5000.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Purification and Polar Localization of Pneumococcal LytB, a Putative Endo-ß-N-Acetylglucosaminidase: the Chain-Dispersing Murein Hydrolase

Blanca De Las Rivas, José L. García, Rubens López,* and Pedro García

Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain

Received 13 March 2002/ Accepted 21 June 2002

The DNA region encoding the mature form of a pneumococcal murein hydrolase (LytB) was cloned and expressed in Escherichia coli. LytB was purified by affinity chromatography, and its activity was suggested to be the first identified endo-ß-N-acetylglucosaminidase of Streptococcus pneumoniae. LytB can remove a maximum of only 25% of the radioactivity from [3H]choline-labeled pneumococcal cell walls in in vitro assays. Inactivation of the lytB gene of wild-type strain R6 (R6B mutant) led to the formation of long chains but did not affect either total cell wall hydrolytic activity at the stationary phase of growth or development of genetic competence. Longer chains were formed when the lytB mutation was introduced into the M31 strain (M31B mutant), which harbors a complete deletion of lytA, which codes for the major autolysin. Furthermore, the use of this mutant revealed that LytB is the first nonautolytic murein hydrolase of pneumococcus. Purified LytB added to pneumococcal cultures of R6B or M31B was capable of dispersing, in a dose-dependent manner, the long chains characteristic of these mutants into diplococci or short chains, the typical morphology of R6 and M31 strains, respectively. In vitro acetylation of purified pneumococcal cell walls did not affect the activity of LytB, whereas that of the LytA amidase was drastically reduced. On the other hand, the use of a translational fusion between the gene (gfp) coding for the green fluorescent protein (GFP) and lytB supports the notion that LytB accumulates in the cell poles of either the wild-type R6, lytB mutants, or ethanolamine-containing cells (EA cells). The GFP-LytB fusion protein was also able to unchain the lytB mutants but not the EA cells. In contrast, translational fusion protein GFP-LytA preferentially bound to the equatorial regions of choline-containing cells but did not affect their average chain length. These observations suggest the existence of specific receptors for LytB that are positioned at the polar region on the pneumococcal surface, allowing localized peptidoglycan hydrolysis and separation of the daughter cells.

* Corresponding author. Mailing address: Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006 Madrid, Spain. Phone: (34)-91 5611800. Fax: (34)-91 5627518. E-mail: ruben{at}cib.csic.es.

Journal of Bacteriology, September 2002, p. 4988-5000, Vol. 184, No. 18
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.18.4988-5000.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Frias, M. J., Melo-Cristino, J., Ramirez, M. (2009). The Autolysin LytA Contributes to Efficient Bacteriophage Progeny Release in Streptococcus pneumoniae. J. Bacteriol. 191: 5428-5440 [Abstract] [Full Text]  
  • Barendt, S. M., Land, A. D., Sham, L.-T., Ng, W.-L., Tsui, H.-C. T., Arnold, R. J., Winkler, M. E. (2009). Influences of Capsule on Cell Shape and Chain Formation of Wild-Type and pcsB Mutants of Serotype 2 Streptococcus pneumoniae. J. Bacteriol. 191: 3024-3040 [Abstract] [Full Text]  
  • Le Gouellec, A., Roux, L., Fadda, D., Massidda, O., Vernet, T., Zapun, A. (2008). Roles of Pneumococcal DivIB in Cell Division. J. Bacteriol. 190: 4501-4511 [Abstract] [Full Text]  
  • Rice, K. C., Bayles, K. W. (2008). Molecular Control of Bacterial Death and Lysis. Microbiol. Mol. Biol. Rev. 72: 85-109 [Abstract] [Full Text]  
  • Attali, C., Frolet, C., Durmort, C., Offant, J., Vernet, T., Di Guilmi, A. M. (2008). Streptococcus pneumoniae Choline-Binding Protein E Interaction with Plasminogen/Plasmin Stimulates Migration across the Extracellular Matrix. Infect. Immun. 76: 466-476 [Abstract] [Full Text]  
  • Dubrac, S., Boneca, I. G., Poupel, O., Msadek, T. (2007). New Insights into the WalK/WalR (YycG/YycF) Essential Signal Transduction Pathway Reveal a Major Role in Controlling Cell Wall Metabolism and Biofilm Formation in Staphylococcus aureus. J. Bacteriol. 189: 8257-8269 [Abstract] [Full Text]  
  • Priyadarshini, R., de Pedro, M. A., Young, K. D. (2007). Role of Peptidoglycan Amidases in the Development and Morphology of the Division Septum in Escherichia coli. J. Bacteriol. 189: 5334-5347 [Abstract] [Full Text]  
  • Mills, M. F., Marquart, M. E., McDaniel, L. S. (2007). Localization of PcsB of Streptococcus pneumoniae and Its Differential Expression in Response to Stress. J. Bacteriol. 189: 4544-4546 [Abstract] [Full Text]  
  • Saskova, L., Novakova, L., Basler, M., Branny, P. (2007). Eukaryotic-Type Serine/Threonine Protein Kinase StkP Is a Global Regulator of Gene Expression in Streptococcus pneumoniae. J. Bacteriol. 189: 4168-4179 [Abstract] [Full Text]  
  • Wang, L., Lin, M. (2007). Identification of IspC, an 86-Kilodalton Protein Target of Humoral Immune Response to Infection with Listeria monocytogenes Serotype 4b, as a Novel Surface Autolysin. J. Bacteriol. 189: 2046-2054 [Abstract] [Full Text]  
  • Fadda, D., Santona, A., D'Ulisse, V., Ghelardini, P., Ennas, M. G., Whalen, M. B., Massidda, O. (2007). Streptococcus pneumoniae DivIVA: Localization and Interactions in a MinCD-Free Context. J. Bacteriol. 189: 1288-1298 [Abstract] [Full Text]  
  • Vicente, M., Garcia-Ovalle, M. (2007). Making a Point: the Role of DivIVA in Streptococcal Polar Anatomy{triangledown}. J. Bacteriol. 189: 1185-1188 [Full Text]  
  • Lanie, J. A., Ng, W.-L., Kazmierczak, K. M., Andrzejewski, T. M., Davidsen, T. M., Wayne, K. J., Tettelin, H., Glass, J. I., Winkler, M. E. (2007). Genome Sequence of Avery's Virulent Serotype 2 Strain D39 of Streptococcus pneumoniae and Comparison with That of Unencapsulated Laboratory Strain R6. J. Bacteriol. 189: 38-51 [Abstract] [Full Text]  
  • Moscoso, M., Garcia, E., Lopez, R. (2006). Biofilm Formation by Streptococcus pneumoniae: Role of Choline, Extracellular DNA, and Capsular Polysaccharide in Microbial Accretion. J. Bacteriol. 188: 7785-7795 [Abstract] [Full Text]  
  • Fukushima, T., Afkham, A., Kurosawa, S.-i., Tanabe, T., Yamamoto, H., Sekiguchi, J. (2006). A New D,L-Endopeptidase Gene Product, YojL (Renamed CwlS), Plays a Role in Cell Separation with LytE and LytF in Bacillus subtilis.. J. Bacteriol. 188: 5541-5550 [Abstract] [Full Text]  
  • Scheffers, D.-J., Pinho, M. G. (2005). Bacterial Cell Wall Synthesis: New Insights from Localization Studies. Microbiol. Mol. Biol. Rev. 69: 585-607 [Abstract] [Full Text]  
  • Moscoso, M., Obregon, V., Lopez, R., Garcia, J. L., Garcia, E. (2005). Allelic Variation of Polymorphic Locus lytB, Encoding a Choline-Binding Protein, from Streptococci of the Mitis Group. Appl. Environ. Microbiol. 71: 8706-8713 [Abstract] [Full Text]  
  • Ng, W.-L., Tsui, H.-C. T., Winkler, M. E. (2005). Regulation of the pspA Virulence Factor and Essential pcsB Murein Biosynthetic Genes by the Phosphorylated VicR (YycF) Response Regulator in Streptococcus pneumoniae. J. Bacteriol. 187: 7444-7459 [Abstract] [Full Text]  
  • Garau, G., Lemaire, D., Vernet, T., Dideberg, O., Di Guilmi, A. M. (2005). Crystal Structure of Phosphorylcholine Esterase Domain of the Virulence Factor Choline-binding Protein E from Streptococcus pneumoniae: NEW STRUCTURAL FEATURES AMONG THE METALLO-{beta}-LACTAMASE SUPERFAMILY. J. Biol. Chem. 280: 28591-28600 [Abstract] [Full Text]  
  • Shibata, Y., Kawada, M., Nakano, Y., Toyoshima, K., Yamashita, Y. (2005). Identification and Characterization of an Autolysin-Encoding Gene of Streptococcus mutans. Infect. Immun. 73: 3512-3520 [Abstract] [Full Text]  
  • Fernandez-Tornero, C., Garcia, E., de Pascual-Teresa, B., Lopez, R., Gimenez-Gallego, G., Romero, A. (2005). Ofloxacin-like Antibiotics Inhibit Pneumococcal Cell Wall-degrading Virulence Factors. J. Biol. Chem. 280: 19948-19957 [Abstract] [Full Text]  
  • Rice, K. C., Nelson, J. B., Patton, T. G., Yang, S.-J., Bayles, K. W. (2005). Acetic Acid Induces Expression of the Staphylococcus aureus cidABC and lrgAB Murein Hydrolase Regulator Operons. J. Bacteriol. 187: 813-821 [Abstract] [Full Text]  
  • Carroll, S. A., Hain, T., Technow, U., Darji, A., Pashalidis, P., Joseph, S. W., Chakraborty, T. (2003). Identification and Characterization of a Peptidoglycan Hydrolase, MurA, of Listeria monocytogenes, a Muramidase Needed for Cell Separation. J. Bacteriol. 185: 6801-6808 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»