Previous Article
J Bacteriol, April 1998, p. 1973-1977, Vol. 180, No. 7
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Characterization of the Aspergillus nidulans
nmrA Gene Involved in Nitrogen Metabolite Repression
Alex
Andrianopoulos,
Sophie
Kourambas,
Julie A.
Sharp,
Meryl A.
Davis, and
Michael
J.
Hynes*
Department of Genetics, University of
Melbourne, Parkville, Victoria 3052, Australia
Received 10 November 1997/Accepted 26 January 1998
 |
ABSTRACT |
The gene nmrA of Aspergillus nidulans has
been isolated and found to be a homolog of the Neurospora
crassa gene nmr-1, involved in nitrogen
metabolite repression. Deletion of nmrA results in partial
derepression of activities subject to nitrogen repression similar to
phenotypes observed for certain mutations in the positively acting
areA gene.
| |
TEXT |
Fungi are capable of using a wide
range of compounds as sources of nitrogen. Genes encoding enzymes and
permeases required for nitrogen utilization are often regulated by
specific induction mechanisms. In addition, they are usually subject to
a general control mechanism, sometimes called nitrogen metabolite
repression, according to which they are expressed at high levels only
under conditions of nitrogen limitation. This enables readily
assimilated nitrogen sources such as ammonium and glutamine to be used
preferentially (for a review, see reference 26).
In all fungi investigated, a key feature of this regulatory mechanism
is activation by regulatory proteins containing a DNA binding domain
consisting of a four-cysteine, single zinc finger characteristic of the
GATA family of transcription factors. In each case, loss-of-function
mutations in the genes encoding these GATA factors result in reduced
ability for growth on many different sole sources of nitrogen.
Saccharomyces cerevisiae has two genes, GLN3 and
NIL1/GAT1, while in other fungi only a single gene has been
found: nit-2 in Neurospora crassa,
areA in Aspergillus nidulans, nut1 in
Magnaportha grisae, and nreA in Penicillium
chrysogenum (4, 6, 17, 18, 21, 25, 27, 34, 36). The
basic model for nitrogen metabolite repression is that growth in the presence of preferred nitrogen sources results in the generation of one
or more signals which antagonize activation of gene expression by GATA
factors. Under nitrogen-limiting conditions, the activators activate
the expression of genes involved in the use of nitrogen sources.
In N. crassa, the gene nmr-1 has been found to be
important for the response to nitrogen metabolite repression. Recessive mutations in this gene result in derepression of some
nitrogen-controlled activities, suggesting a negative role for the gene
(16, 31, 32, 37). Cloning and characterization of this gene
have enabled studies of its role in nitrogen metabolite repression
(19, 23, 24, 40). NIT2 and NMR1 were shown to interact by
the use of the yeast two-hybrid system as well by in vitro assays. Two
regions of NIT2 have been shown to be involved in interactions with
NMR1, one in the conserved region adjacent to the zinc finger and one consisting of the 12 carboxyl-terminal residues. Mutations in both of
these regions prevent interaction with NMR1 and result in derepressed
phenotypes (39). These data strongly suggest that at least
one component of nitrogen metabolite repression in N. crassa
involves NMR1 interacting with NIT2 under nitrogen-sufficient conditions to prevent NIT2 binding to its recognition sequences and
activating gene expression. Some in vitro binding studies support this
model (39).
Extensive mutagenesis studies of areA in A. nidulans have shown that residues within the AreA DNA binding
domain as well as in the carboxyl-terminal region lead to some degree
of derepression for nitrogen-regulated activities (25, 29, 30,
35). In addition to the DNA binding domains, the carboxyl termini
of NIT2, NreA and AreA, are highly conserved (29). Further,
it has been shown that the deletion of sequences within the 3'
untranslated region of areA results in some derepression and
that this correlates with a stabilization of areA mRNA in
ammonium-grown mycelia (29). The sequences involved are
conserved in the P. chrysogenum nreA homolog. The gene
nit-2 has been shown to complement areA
loss-of-function mutations in A. nidulans (12).
However, partial derepression of the various nitrogen-regulated
activities was observed. The nit-2 plasmid used may have
lacked the necessary 3' untranslated sequences (13). The
xprD1 mutation, isolated as leading to derepression of protease expression (7), is an inversion truncating
areA such that the 3' coding and untranslated regions of the
areA mRNA are missing (2, 25). Truncation of
areA as in the xprD1 mutation would result in the
loss of both mechanisms; this has been confirmed by the construction of
appropriate double areA deletion mutations (29).
Therefore there is strong evidence for two distinct mechanisms of
modulation for areA and nit-2 activity,
protein-protein interactions affecting DNA binding and nitrogen
regulation of mRNA stability.
Although no mutants with the predicted phenotype have been isolated,
the data lead to the strong prediction that A. nidulans has a homolog of nmr-1 of N. crassa. Expression of nmr-1 in A. nidulans
suggests that this is the case (Polley and Caddick as cited in
reference 26). We have confirmed this by cloning the nmr-1 homolog from A. nidulans and have found a
central extended conserved region. Disruption of the gene results
in partially derepressed phenotypes similar to those observed in
areA mutants with alterations in the DNA binding domain and
the carboxyl terminus.
Cloning and analysis of nmrA.
A search of the A. nidulans expressed sequence tag (EST) database (32a)
with the N. crassa nmr-1 predicted protein sequence (accession no. P23762) revealed a sequence encoding a
polypeptide fragment with extensive similarity. This allowed the design
of primers for amplification from A. nidulans genomic
DNA of a 383-bp sequence by PCR (Fig.
1A). Cloning and
sequencing of this fragment confirmed homology with
nmr-1 and the presence of a 68-bp intron. Southern blot
analysis of A. nidulans genomic DNA indicated that the
amplified sequence was unique and allowed the identification of a 7-kb
XbaI-BglII hybridizing fragment. This fragment
was cloned into XbaI-BamHI-digested pBLUESCRIPT
SK+ (Stratagene, Inc.) by generating a partial genomic library and
probing colony lifts with the PCR fragment. Restriction
mapping confirmed that the arrangements of sites within the genome and
the clone were identical (Fig. 1A).
View larger version (34K):
[in this window]
[in a new window]
|
FIG. 1.
(A) Restriction map of the 7-kb
XbaI-BglII fragment containing the gene
nmrA. The two exons of the coding region (open rectangles)
and the direction of transcription (arrow) are shown. The flanking
coordinates of the open reading frame relative to the translational
start are shown below the map. The location and coordinates of the EST
(hatched rectangle) are shown. The gene nmrA was sequenced
with a combination of dye primer and dye terminator reactions on an ABI
377 sequencer by the strategy shown, with the direction and size of the
arrows indicating the strand and length of sequence. Restriction sites
are BamHI (B), BglII (Bg), ClaI
(C), EcoRI (E), EcoRV (V), HindIII
(H), KpnI (K), PstI (P),
SacI (Sc), SalI (S), SmaI (Sm), and
XbaI (X). (B) Deletion constructs. pALX180, in which the
EcoRV fragment spanning the entire coding region of
nmrA as well as approximately 800 and 200 bp of 5' and 3'
sequences, respectively, was replaced with an end-filled
BamHI fragment containing the bleomycin resistance gene from
pAmPh520 (3), and pALX183, in which the
ClaI-EcoRV fragment spanning the entire coding
region of nmrA as well as approximately 800 bp from both 5'
and 3' sequences was replaced with a ClaI-SmaI
fragment containing the A. nidulans gene argB
(38). The flanking sequences (solid line) and selectable
markers (open rectangles) and their direction of transcription (arrow)
are shown. (C) Nucleotide and conceptual protein sequence of
nmrA. The region encompassed by the EST (underlined) and the
oligonucleotide primers NMRA and NMRB (arrow) used for the
PCR-generated nmrA probe are shown. The two EcoRV
restriction sites are marked for reference to the map in panel A. Nucleotides are numbered with reference to the +1 at the start of the
coding region.
|
|
Sequence determination of 2,173 bp flanking the PCR fragment indicated
an open reading frame encoding a 352-amino-acid polypeptide
which is
interrupted by a single intron (Fig.
1C). Comparison
of the predicted
polypeptide with that of
N. crassa nmr-1 showed
similarity extending throughout the sequence, with five regions
of very
high conservation (Fig.
2). However, the
N. crassa sequence
is extended at both the amino and
carboxyl termini by 59 and 77
amino acids, respectively. It has been
previously shown that the
introduction of stop codons into
nmr-1, leading to the loss of
up to 77 carboxyl-terminal
amino acids, does not appear to affect
function, while function is
abolished by the loss of 104 carboxyl-terminal
amino acids
(
40). The former truncation removes all additional
carboxyl-terminal amino acids of NMR1, while the latter removes
the
fifth highly conserved region between NMR1 and NmrA. In addition,
protein-protein interaction studies have shown that the 45 amino-terminal
amino acids of NMR1, not present in NmrA, are not
required for
binding to NIT2 and that the region from amino acids
118 to 284,
spanning the three central conserved regions
between NMR1 and
NmrA, also binds NIT2, although the interaction
is weaker (
39).
One insertion of 26 residues in NMR1 is not
present in NmrA, and
one insertion of 28 residues in NmrA is not
present in NMR1. Inspection
of the DNA sequences encoding these
insertions indicate that they
could have arisen by the mutation of
intron splice sites (Fig.
1C) (
40).
View larger version (75K):
[in this window]
[in a new window]
|
FIG. 2.
Alignment of the A. nidulans NmrA and
N. crassa NMR1 conceptual protein sequences. The alignment
was generated with GAP from the Wisconsin package (version 8; Genetics
Computer Group, Madison, Wis.) with the Dayhoff protein comparison
weight matrix, a GAP weight of 3, and a length weight of 0.1. Identities between the two sequences are marked with vertical lines,
and similarities are marked by colons. The five regions of highest
identity are shaded. The two sequences show 60.8% identity and 75.5%
similarity over their entire region.
|
|
Characterization of nmrA deletion strains.
Two
different constructs were used to create nmrA deletions. In
pALX180 the bleomycin resistance gene from Tn5 expressed
from the N. crassa am promoter (3) replaced
the nmrA sequence (Fig. 1). A linear
SacI-KpnI fragment containing the
nmrA::bleoR insert was used
to transform A. nidulans MH3408 (biA1
amdS::lacZ niiA4), selecting for
resistance to bleomycin. Approximately 20% of transformants showed
some phenotypes characteristic of derepression of nitrogen-regulated
activities (see below). In pALX183, argB (38)
replaced the nmrA sequence (Fig. 1), and a
NotI-KpnI fragment containing the
nmrA::argB insert was used to transform
A. nidulans MH8826 (yA1 pabaA1 argB1
amdA7), selecting for arginine prototrophy. Approximately
20% of transformants showed derepression phenotypes. Southern
blot analysis of DNA isolated from transformants confirmed that,
for each construct, the observed phenotypes correlated with replacement of nmrA sequences with
bleoR or argB+
gene, respectively. One transformant from a single nmrA
deletion event was isolated from each experiment and used for further
characterization.
Various plate tests can be used to determine derepression of activities
subject to nitrogen metabolite repression (reference
29 and references therein). The
nmrA
deletion transformants
as well as an
xprD1-containing strain
were sensitive to the toxic
effects of aspartylhydroxamate in the
presence of ammonium, an
indication of derepression of asparaginase
(
15) (Fig.
3). The
nmrA deletion also resulted in slight sensitivity to
thiourea
in the presence of ammonium, but the sensitivity was not as
great
as that of the
xprD1-containing strain (Fig.
3).
Thiourea toxicity
is an indicator of the activity of the
ureA-encoded urea permease
(
28). Similarly,
sensitivity to chlorate, a toxic analog of
nitrate, in the presence of
ammonium was observed in the
nmrA deletion strain, but the
sensitivity was intermediate between
the
xprD1 and wild-type
strains (Fig.
3). Tests for derepression
of extracellular protease
production by the observation of a halo
of clearing of milk (0.5 to
1.0%) in the presence of ammonium
indicated that the
nmrA
deletion strains, unlike
xprD1-containing
strains, were not
detectably derepressed. These phenotypes were
similar to those observed
by Platt et al. (29) for
areA mutant
strains encoding AreA
proteins truncated at the carboxyl-terminal
end but encoding mRNA with
an intact 3' untranslated region and
indicated partial derepression for
some activities.
View larger version (68K):
[in this window]
[in a new window]
|
FIG. 3.
Growth properties of nmrA deletion strains.
Growth was scored for 2 to 3 days at 37°C on 1% glucose medium
(9) containing 5 mM ammonium tartrate together with 200 mM
potassium chlorate (ClO3), 5 mM ammonium tartrate together
with 5 mM D,L- -aspartylhydroxamate (AH), and
2.5 mM ammonium tartrate with 10 mM thiourea (TU) with appropriate
auxotrophic supplements. The
 nmrA::argB+ strain was
generated by transformation of a strain whose genotype was yA1
pabaA1 argB1 amdA7 with a gel-purified insert of pALX183 selecting
for arginine prototrophy (Fig. 1B). The nmrA+
(argB+) control strain was an ArgB+
transformant from the same transformation that did not result in the
deletion of nmrA. The
nmrA::bleoR strain was
obtained by transformation of a strain whose genotype was biA1
amdS::lacZ niiA4 with a gel-purified insert
of pALX180 (Fig. 1B) selecting for resistance to bleomycin. The
nmrA+ (bleoR) control
strain was a bleomycin-resistant transformant from the same
transformation that did not result in the deletion of nmrA.
The genotypes of the strains designated wild type and xprD1
were biA1 and biA1 amdS::lacZ
xprD1 niiA4, respectively. Genotypes and phenotypes were reported
by Clutterbuck (5). The xprD1 mutation results
from an inversion truncating the areA gene (2,
25).
|
|
The
nmrA::
argB deletion strain
was transformed with the bleomycin resistance-encoding
plasmid pAmPh520 (
3) together with
a plasmid (pALX186)
containing
nmrA on an
EcoRV fragment (Fig.
1) and selecting for bleomycin resistance. Approximately 50% of
the
transformants were found to be phenotypically
nmrA+ as shown by resistance to
aspartylhydroxamate and chlorate in
the presence of ammonium. This
finding indicated that these phenotypes
resulted from the deletion of
nmrA and that this fragment is sufficient
for
nmrA function.
The gene
amdS, encoding acetamidase, is subject to nitrogen
metabolite repression (
14). The deletion of
nmrA
was found to
result in partial derepression of the expression of an
amdS::
lacZ reporter gene
(
11) with respect to both ammonium and glutamine
(Table
1). Partial derepression was also
observed in the presence
of GABA as inducer, reflecting derepression of
both
amdS expression
and GABA uptake via the
gabA-encoded permease (
1). Partial
depression
for nitrate reductase was also observed (Table
2),
and the level of derepression was
similar to that reported for
the
areA mutants encoding
proteins truncated at the carboxyl terminus
(
29,
35).
The
areA102 mutation, resulting from an amino acid
substitution in the zinc finger region, is a change in specificity
mutation
resulting in increased activation of some nitrogen-controlled
activities and decreased activation of others (
22,
25). A
cross between the
nmrA::
bleoR deletion strain
and an
areA102 strain resulted in
areA102
nmrA::
bleoR double
mutants which showed increased sensitivity to chlorate
and
thiourea in the presence of ammonium relative to the
nmrA::
bleoR single
mutant and derepression for extracellular protease activity
as shown by
a halo of milk clearing in the presence of either
ammonium or
glutamine.
These results clearly indicate that one of the mechanisms for nitrogen
metabolite repression is conserved between
N. crassa and
A. nidulans, namely protein-protein interactions between
NMR1/NmrA
and NIT2/AreA in the presence of sources of repression. The
magnitude
of the effects of deletion of
nmrA on nitrogen
metabolite repression
is similar to that observed for deletion of the
conserved 12 carboxyl-terminal
amino acids of AreA (
29). It
is predicted that the effects of
these mutations will not be additive
in double mutants. Since
deletion of
nmrA does not result in
complete derepression, this
gene is unlikely to be involved in
modulation of
areA mRNA stability
via sequences in the 3'
untranslated region (
29). Therefore,
it is
predicted that double mutants containing an
nmrA
deletion
and a deletion of the 3' untranslated region of
areA will show
additive levels of derepression, as
previously observed for the
xprD1 inversion mutation
and for
areA mutants with deletions of
both the
carboxyl terminus and the relevant 3' untranslated sequences
(
29).
The nature of the signal or signals generated by nitrogen metabolites
and the question of whether both mechanisms have any
common components
remain to be determined. Furthermore, the role
of negatively acting
factors of the GATA family as found in
S. cerevisiae
(
8,
10,
33), and recently suggested to occur
in filamentous
fungi (
20), needs to be investigated.
Nucleotide sequence accession number.
The sequence for the
nmrA gene has been deposited in GenBank under accession no.
AF041976.
| |
ACKNOWLEDGMENTS |
This work was supported by a grant from the Australian Research
Council. Database and computer analysis was performed with the
Australian National Genomics Information Service (ANGIS).
Assistance by Kathleen Soltys is greatly appreciated.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Department of
Genetics, University of Melbourne, Parkville, Victoria 3052, Australia. Phone: 61 3 9344 5140. Fax: 61 3 9344 5139. E-mail:
hynes.lab{at}genetics.unimelb.edu.au.
| |
REFERENCES |
| 1.
|
Arst, H. N.
1976.
Integrator gene in Aspergillus nidulans.
Nature
262:231-234[Medline].
|
| 2.
|
Arst, H. N.
1982.
A near terminal pericentric inversion leads to nitrogen metabolite derepression in Aspergillus nidulans.
Mol. Gen. Genet.
188:490-493[Medline].
|
| 3.
|
Austin, B.,
R. M. Hall, and B. M. Tyler.
1990.
Optimized vectors and selection for transformation of Neurospora crassa and Aspergillus nidulans to bleomycin and phleomycin resistance.
Gene
93:157-162[Medline].
|
| 4.
|
Caddick, M. X.,
H. N. Arst,
L. H. Taylor,
R. I. Johnson, and A. G. Brownlee.
1986.
Cloning of the regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans.
EMBO J.
5:1087-1090[Medline].
|
| 5.
|
Clutterbuck, A. J.
1974.
Aspergillus nidulans genetics, p. 447-510. In
R. C. King (ed.), Handbook of genetics, vol. 1.
Plenum Publishing Corp., New York, N.Y.
|
| 6.
|
Coffman, J. A.,
R. Rai,
T. Cunningham,
V. Svetlov, and T. G. Cooper.
1996.
Gat1p, a GATA family protein whose production is sensitive to nitrogen catabolite repression, participates in transcriptional activation of nitrogen-catabolic genes in Saccharomyces cerevisiae.
Mol. Cell. Biol.
16:847-858[Abstract].
|
| 7.
|
Cohen, B. L.
1972.
Ammonium repression of extracellular proteases in Aspergillus nidulans.
J. Gen. Microbiol.
71:293-299.
|
| 8.
|
Coornaert, D.,
S. Vissers,
B. Andre, and M. Grenson.
1992.
The UGA43 negative regulatory gene of Saccharomyces cerevisiae contains both a GATA-1 type zinc finger and a putative leucine zipper.
Curr. Genet.
21:301-307[Medline].
|
| 9.
|
Cove, D. J.
1966.
The induction and repression of nitrate reductase in the fungus Aspergillus nidulans.
Biochim. Biophys. Acta
113:51-56[Medline].
|
| 10.
|
Cunningham, T. S., and T. G. Cooper.
1991.
Expression of the DAL80 gene, whose product is homologous to the GATA factors and is a negative regulator of multiple nitrogen catabolic genes in Saccharomyces cerevisiae, is sensitive to nitrogen catabolite repression.
Mol. Cell. Biol.
11:6205-6215[Abstract/Free Full Text].
|
| 11.
|
Davis, M. A.,
C. S. Cobbett, and M. J. Hynes.
1988.
An amdS-lacZ fusion for studying gene regulation in Aspergillus.
Gene
63:199-212[Medline].
|
| 12.
|
Davis, M. A., and M. J. Hynes.
1987.
Complementation of areA regulatory gene mutations of Aspergillus nidulans by the heterologous regulatory gene nit-2 of Neurospora crassa.
Proc. Natl. Acad. Sci. USA
84:3753-3757[Abstract/Free Full Text].
|
| 13.
|
Davis, M. A., and M. J. Hynes.
1997.
Why are nit-2 transformants of Aspergillus nidulans partially derepressed?
Fungal Genet. Newsl.
44:13-14.
|
| 14.
|
Davis, M. A.,
J. M. Kelly, and M. J. Hynes.
1993.
Fungal catabolic gene regulation: molecular genetic analysis of the amdS gene of Aspergillus nidulans.
Genetica
90:133-145[Medline].
|
| 15.
|
Drainas, C., and J. A. Pateman.
1977.
L-Asparaginase activity in the fungus Aspergillus nidulans.
Biochem. Soc. Trans.
5:259-261[Medline].
|
| 16.
|
Dunn-Coleman, N. S.,
A. B. Tomsett, and R. H. Garrett.
1981.
The regulation of nitrate assimilation in Neurospora crassa: biochemical analysis of the nmr-1 mutants.
Mol. Gen. Genet.
182:234-239[Medline].
|
| 17.
|
Froeliger, E. H., and B. E. Carpenter.
1996.
NUT1, a major nitrogen regulatory gene in Magnaporthe grisea, is dispensable for pathogenicity.
Mol. Gen. Genet.
251:647-656[Medline].
|
| 18.
|
Fu, Y. H., and G. A. Marzluf.
1987.
Molecular cloning and analysis of the regulation of nit-3, the structural gene for nitrate reductase in Neurospora crassa.
Proc. Natl. Acad. Sci. USA
84:8243-8247[Abstract/Free Full Text].
|
| 19.
|
Fu, Y. H.,
J. L. Young, and G. A. Marzluf.
1988.
Molecular cloning and characterization of a negative-acting nitrogen regulatory gene of Neurospora crassa.
Mol. Gen. Genet.
214:74-79[Medline].
|
| 20.
|
Haas, H.,
K. Angermayr,
I. Zadra, and G. Stoffler.
1997.
Overexpression of nreB, a new gata factor-encoding gene of Penicillium chrysogenum, leads to repression of the nitrate assimilatory gene cluster.
J. Biol. Chem.
272:22576-22582[Abstract/Free Full Text].
|
| 21.
|
Haas, H.,
B. Bauer,
B. Redl,
G. Stoffler, and G. A. Marzluf.
1995.
Molecular cloning and analysis of nre, the major nitrogen regulatory gene of Penicillium chrysogenum.
Curr. Genet.
27:150-158[Medline].
|
| 22.
|
Hynes, M. J.
1975.
Studies on the role of the areA gene in the regulation of nitrogen catabolism in Aspergillus nidulans.
Aust. J. Biol. Sci.
28:301-313[Medline].
|
| 23.
|
Jarai, G., and G. A. Marzluf.
1990.
Analysis of conventional and in vitro generated mutants of nmr, the negatively acting nitrogen regulatory gene of Neurospora crassa.
Mol. Gen. Genet.
222:233-420[Medline].
|
| 24.
|
Jarai, G., and G. A. Marzluf.
1991.
Generation of new mutants of nmr, the negative-acting nitrogen regulatory gene of Neurospora crassa, by repeat induced mutation.
Curr. Genet.
20:283-288[Medline].
|
| 25.
|
Kudla, B.,
M. X. Caddick,
T. Langdon,
N. M. Martinez-Rossi,
C. F. Bennett,
S. Sibley,
R. W. Davies, and H. N. Arst.
1990.
The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger.
EMBO J.
9:1355-1364[Medline].
|
| 26.
|
Marzluf, G. A.
1997.
Genetic regulation of nitrogen metabolism in the fungi.
Microbiol. Mol. Biol. Rev.
61:17-32.
[Abstract] |
| 27.
|
Minehart, P. L., and B. Magasanik.
1991.
Sequence and expression of GLN3, a positive nitrogen regulatory gene of Saccharomyces cerevisiae encoding a protein with a putative zinc finger DNA-binding domain.
Mol. Cell. Biol.
11:6216-6228[Abstract/Free Full Text].
|
| 28.
|
Pateman, J. A.,
E. Dunn, and E. M. Mackay.
1982.
Urea and thiourea transport in Aspergillus nidulans.
Biochem. Genet.
20:777-790[Medline].
|
| 29.
|
Platt, A.,
T. Langdon,
H. N. Arst,
D. Kirk,
D. Tollervey,
J. M. Sanchez, and M. X. Caddick.
1996.
Nitrogen metabolite signalling involves the C-terminus and the GATA domain of the Aspergillus transcription factor AREA and the 3' untranslated region of its mRNA.
EMBO J.
15:2791-2801[Medline].
|
| 30.
|
Platt, A.,
A. Ravagnani,
H. N. Arst,
D. Kirk,
T. Langdon, and M. X. Caddick.
1996.
Mutational analysis of the C-terminal region of AREA, the transcription factor mediating nitrogen metabolite repression in Aspergillus nidulans.
Mol. Gen. Genet.
250:106-114[Medline].
|
| 31.
|
Premakumar, R.,
G. J. Sorger, and D. Gooden.
1979.
Nitrogen metabolite repression of nitrate reductase in Neurospora crassa.
J. Bacteriol.
137:1119-1126[Abstract/Free Full Text].
|
| 32.
|
Premakumar, R.,
G. J. Sorger, and D. Gooden.
1980.
Physiological characterization of a Neurospora crassa mutant with impaired regulation of nitrate reductase.
J. Bacteriol.
144:542-551[Abstract/Free Full Text].
|
| 32a.
| Roe, B. A., D. Kupfer, S. Clifton, and R. A. Prade. Aspergillus nidulans cDNA Sequencing Project. URL
http://www.genome.ou.edu/asper.html.
|
| 33.
|
Soussi-Boudekou, S.,
S. Vissers,
A. Urrestarazu,
J. C. Jauniaux, and B. Andre.
1997.
Gzf3p, a fourth GATA factor involved in nitrogen-regulated transcription in Saccharomyces cerevisiae.
Mol. Microbiol.
23:1157-1168[Medline].
|
| 34.
|
Stanbrough, M.,
D. W. Rowen, and B. Magasanik.
1995.
Role of the GATA factors Gln3p and Nil1p of Saccharomyces cerevisiae in the expression of nitrogen-regulated genes.
Proc. Natl. Acad. Sci. USA
92:9450-9454[Abstract/Free Full Text].
|
| 35.
|
Stankovich, M.,
A. Platt,
M. X. Caddick,
T. Langdon,
P. M. Shaffer, and H. N. Arst.
1993.
C-terminal truncation of the transcriptional activator encoded by areA in Aspergillus nidulans results in both loss-of-function and gain-of-function phenotypes.
Mol. Microbiol.
7:81-87[Medline].
|
| 36.
|
Stewart, V., and S. J. Vollmer.
1986.
Molecular cloning of nit-2, a regulatory gene required for nitrogen metabolite repression in Neurospora crassa.
Gene
46:291-295[Medline].
|
| 37.
|
Tomsett, A. B.,
N. S. Dunn-Coleman, and R. H. Garrett.
1981.
The regulation of nitrate assimilation in Neurospora crassa: the isolation and genetic analysis of nmr-1 mutants.
Mol. Gen. Genet.
182:229-233[Medline].
|
| 38.
|
Upshall, A.,
T. Gilbert,
G. Saari,
P. J. O'Hara,
P. Weglenski,
B. Berse,
K. Miller, and W. E. Timberlake.
1986.
Molecular analysis of the argB gene of Aspergillus nidulans.
Mol. Gen. Genet.
204:349-354[Medline].
|
| 39.
|
Xiao, X.,
Y. H. Fu, and G. A. Marzluf.
1995.
The negative-acting NMR regulatory protein of Neurospora crassa binds to and inhibits the DNA-binding activity of the positive-acting nitrogen regulatory protein NIT2.
Biochemistry
34:8861-8868[Medline].
|
| 40.
|
Young, J. L.,
G. Jarai,
Y. H. Fu, and G. A. Marzluf.
1990.
Nucleotide sequence and analysis of NMR, a negative-acting regulatory gene in the nitrogen circuit of Neurospora crassa.
Mol. Gen. Genet.
222:120-128[Medline].
|
J Bacteriol, April 1998, p. 1973-1977, Vol. 180, No. 7
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Jiang, N., Xiao, D., Zhang, D., Sun, N., Yan, B., Zhu, X.
(2009). Negative Roles of a Novel Nitrogen Metabolite Repression-Related Gene, TAR1, in Laccase Production and Nitrate Utilization by the Basidiomycete Cryptococcus neoformans. Appl. Environ. Microbiol.
75: 6777-6782
[Abstract]
[Full Text]
-
Schonig, B., Brown, D. W., Oeser, B., Tudzynski, B.
(2008). Cross-Species Hybridization with Fusarium verticillioides Microarrays Reveals New Insights into Fusarium fujikuroi Nitrogen Regulation and the Role of AreA and NMR. Eukaryot Cell
7: 1831-1846
[Abstract]
[Full Text]
-
Wong, K. H., Hynes, M. J., Davis, M. A.
(2008). Recent Advances in Nitrogen Regulation: a Comparison between Saccharomyces cerevisiae and Filamentous Fungi. Eukaryot Cell
7: 917-925
[Full Text]
-
Rolland, S. G., Bruel, C. A.
(2008). Sulphur and nitrogen regulation of the protease-encoding ACP1 gene in the fungus Botrytis cinerea: correlation with a phospholipase D activity. Microbiology
154: 1464-1473
[Abstract]
[Full Text]
-
Bernreiter, A., Ramon, A., Fernandez-Martinez, J., Berger, H., Araujo-Bazan, L., Espeso, E. A., Pachlinger, R., Gallmetzer, A., Anderl, I., Scazzocchio, C., Strauss, J.
(2007). Nuclear Export of the Transcription Factor NirA Is a Regulatory Checkpoint for Nitrate Induction in Aspergillus nidulans. Mol. Cell. Biol.
27: 791-802
[Abstract]
[Full Text]
-
Teichert, S., Wottawa, M., Schonig, B., Tudzynski, B.
(2006). Role of the Fusarium fujikuroi TOR Kinase in Nitrogen Regulation and Secondary Metabolism.. Eukaryot Cell
5: 1807-1819
[Abstract]
[Full Text]
-
Monahan, B. J., Askin, M. C., Hynes, M. J., Davis, M. A.
(2006). Differential Expression of Aspergillus nidulans Ammonium Permease Genes Is Regulated by GATA Transcription Factor AreA. Eukaryot Cell
5: 226-237
[Abstract]
[Full Text]
-
Todd, R. B., Fraser, J. A., Wong, K. H., Davis, M. A., Hynes, M. J.
(2005). Nuclear Accumulation of the GATA Factor AreA in Response to Complete Nitrogen Starvation by Regulation of Nuclear Export. Eukaryot Cell
4: 1646-1653
[Abstract]
[Full Text]
-
Fitzgibbon, G. J., Morozov, I. Y., Jones, M. G., Caddick, M. X.
(2005). Genetic Analysis of the TOR Pathway in Aspergillus nidulans. Eukaryot Cell
4: 1595-1598
[Abstract]
[Full Text]
-
Lamb, H. K., Leslie, K., Dodds, A. L., Nutley, M., Cooper, A., Johnson, C., Thompson, P., Stammers, D. K., Hawkins, A. R.
(2003). The Negative Transcriptional Regulator NmrA Discriminates between Oxidized and Reduced Dinucleotides. J. Biol. Chem.
278: 32107-32114
[Abstract]
[Full Text]
-
Fraser, J. A., Davis, M. A., Hynes, M. J.
(2002). A Gene from Aspergillus nidulans with Similarity to URE2 of Saccharomyces cerevisiae Encodes a Glutathione S-Transferase Which Contributes to Heavy Metal and Xenobiotic Resistance. Appl. Environ. Microbiol.
68: 2802-2808
[Abstract]
[Full Text]
-
Margelis, S., D'Souza, C., Small, A. J., Hynes, M. J., Adams, T. H., Davis, M. A.
(2001). Role of Glutamine Synthetase in Nitrogen Metabolite Repression in Aspergillus nidulans. J. Bacteriol.
183: 5826-5833
[Abstract]
[Full Text]
-
Hajjaj, H., Niederberger, P., Duboc, P.
(2001). Lovastatin Biosynthesis by Aspergillus terreus in a Chemically Defined Medium. Appl. Environ. Microbiol.
67: 2596-2602
[Abstract]
[Full Text]
-
Fraser, J. A., Davis, M. A., Hynes, M. J.
(2001). The Formamidase Gene of Aspergillus nidulans: Regulation by Nitrogen Metabolite Repression and Transcriptional Interference by an Overlapping Upstream Gene. Genetics
157: 119-131
[Abstract]
[Full Text]
-
Small, A. J., Hynes, M. J., Davis, M. A.
(1999). The TamA Protein Fused to a DNA-Binding Domain Can Recruit AreA, the Major Nitrogen Regulatory Protein, to Activate Gene Expression in Aspergillus nidulans. Genetics
153: 95-105
[Abstract]
[Full Text]
-
Caddick, M. X., Arst, H. N. Jr.
(1998). Deletion of the 389 N-Terminal Residues of the Transcriptional Activator AREA Does Not Result in Nitrogen Metabolite Derepression in Aspergillus nidulans. J. Bacteriol.
180: 5762-5764
[Abstract]
[Full Text]
-
Wilson, R. A., Arst, H. N. Jr.
(1998). Mutational Analysis of AREA, a Transcriptional Activator Mediating Nitrogen Metabolite Repression in Aspergillus nidulans and a Member of the "Streetwise" GATA Family of Transcription Factors. Microbiol. Mol. Biol. Rev.
62: 586-596
[Abstract]
[Full Text]
Top
Abstract
Text
