Conserved domains in bacterial regulatory proteins that respond to environmental stimuli

Ronson, Clive W.; Nixon, B. Tracy; Ausubel, Frederick M.

doi:10.1016/0092-8674(87)90530-7

Cited by 468 publications

(290 citation statements)

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“…4) which are now thought to act in this way (7). The phosphorylation of regulator proteins provides a mechanism for cross-talk (40), in which different genetic and physiological regulatory systems may interact. Cross-talk is almost certainly responsible for the transcriptional regulation of the PHO regulon in phoR mutants.…”

Section: Resultsmentioning

confidence: 99%

Identification of phosphate starvation-inducible genes in Escherichia coli K-12 by DNA sequence analysis of psi::lacZ(Mu d1) transcriptional fusions

1990

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Twenty-four independent phosphate starvation-inducible (psi) transcriptional fusions made with Mu dl(lacZ bla) were analyzed by sequencing the psi::lacZ(Mu dl) chromosomal junctions by using DNAs amplified with the polymerase chain reaction or mini-Mu cloning. Our DNA sequence analysis showed that the MuR DNA in Mu dl has an unexpected structure that is comprised of 104 bases of MuR DNA in the form of a large inverted repeat, which we denoted Mu dl-R. Also, Mu dls in the phoA and phn (psiD) loci of the phosphate regulon showed regional specificities for the insertion sites despite the randomness of Mu dl insertions into the genome as a whole. Gene products or open reading frames were identified for seven unknown psi::lacZ(Mu dl) transcriptional fusions by searching DNA data bases with the sequences adjacent and upstream of the Mu dls. One psiC::lacZ(Mu dl) lies in the ugpB gene of the ugpBAEC operon, which encodes a periplasmic sn-glycerol-3-phosphate-binding protein; two psiQ::lacZ(Mu dl)s lie in the gltB gene, and one psiQ::lacZ(Mu dl) lies in the gltD gene of the gltBDF operon, encoding the large and small subunits of glutamate synthase, respectively; and the psi-51::lacZ(Mu dl) lies in the glpB gene of the glpABC operon, which codes for the anaerobically regulated glycerol-3-phosphate dehydrogenase. psiE and psiF::lacZ(Mu dl)s lie in uncharacterized open reading frames near the xyLE and phoA genes, respectively. Six other psi::lacZ(Mu dl)s lie in yet unreported Escherichia coli sequences.Transcriptional regulation is one of the most intensively studied topics in modern biology. These studies rely upon a number of methods for detection and quantitation of the protein products of a gene or operon. The development of transcriptional and gene (protein) fusion technology has proved valuable in the study of genetic regulatory systems in novel ways, especially in cases where the gene products are difficult to assay directly or are themselves unknown. In such cases, gene regulation can be studied by fusing a reporter gene, minus its own regulatory sequences, to the gene or promoter of interest. The most commonly used reporter gene is lacZ, which encodes 3-galactosidase, for it can be easily assayed. Although numerous techniques for making lacZ transcriptional and translational fusions now exist, one of the more common methods employed in gramnegative bacteria involves in vivo construction with transposons like Mu dl (8).Mu dl is a defective Mu-1 bacteriophage that, like Mu-i, transposes somewhat randomly into Escherichia coli chromosomal or plasmid DNAs (8). The MuR and MuL ends of Mu DNA and the Mu A and B genes are required for transposition, although the Mu A and B products can be supplied in trans (10). In Mu dl the lacZ reporter gene is near MuR so that insertion of Mu dl into a gene in the correct orientation places the lacZ gene under the control of the regulatory sequences of the interrupted gene. The precise physical structure of the MuR end in Mu dl was not determined.Mu dl was used to study Pi control of ...

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Section: Resultsmentioning

confidence: 99%

Identification of phosphate starvation-inducible genes in Escherichia coli K-12 by DNA sequence analysis of psi::lacZ(Mu d1) transcriptional fusions

1990

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“…Membranes were blocked by incubation in Tris-buffered saline with Tween 20 (TBST [37]) plus 5% dried milk (Carnation). The primary antibody was a polyclonal rabbit antisera raised against purified Spo0A tagged with six histidines.…”

Section: Methodsmentioning

confidence: 99%

Control of Sporulation Gene Expression in Bacillus subtilis by the Chromosome Partitioning Proteins Soj (ParA) and Spo0J (ParB)

Quisel

Grossman

2000

J Bacteriol

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Two chromosome partitioning proteins, Soj (ParA) and Spo0J (ParB), regulate the initiation of sporulation in Bacillus subtilis. In a spo0J null mutant, sporulation is inhibited by the action of Soj. Soj negatively regulates expression of several sporulation genes by binding to the promoter regions and inhibiting transcription. All of the genes known to be inhibited by Soj are also activated by the phosphorylated form of the transcription factor Spo0A (Spo0AϳP). We found that, in a spo0J null mutant, Soj affected sporulation, in part, by decreasing the level of Spo0A protein. Soj negatively regulated transcription of spo0A and associated with the spo0A promoter region in vivo. Expression of spo0A from a heterologous promoter in a spo0J null mutant restored Spo0A levels and partly bypassed the sporulation and gene expression defects. Soj did not appear to significantly affect phosphorylation of Spo0A. Thus, in the absence of Spo0J, Soj inhibits sporulation and sporulation gene expression by inhibiting accumulation of the activator protein Spo0A and by acting downstream of Spo0A to inhibit gene expression directly.Bacillus subtilis can sporulate in response to starvation and high cell density. Sporulation involves an asymmetric cell division and requires DNA replication and chromosome segregation. Spo0A is a critical transcription factor in early sporulation. Cells initiate sporulation when they accumulate a threshold amount of phosphorylated Spo0A (Spo0AϳP) (3,6,12). Many signals that regulate sporulation do so by affecting the accumulation of Spo0AϳP. Spo0A obtains phosphate from a set of proteins called the phosphorelay (2). A balance of kinases and phosphatases regulates phosphate input into the phosphorelay and the phosphorylation state of Spo0A (3,(30)(31)(32).Two proteins involved in chromosome partitioning, Soj and Spo0J, also regulate the initiation of sporulation, perhaps in response to chromosome structure or partitioning (4,14,28). Soj and Spo0J are members of the ParA-SopA and ParB-SopB protein families, respectively. Many ParA-and ParB-type proteins are encoded on low-copy-number plasmids and are required for accurate plasmid partitioning (29, 42). Chromosome-encoded family members have also been identified, several of which function in chromosome partitioning (14,22,26). Members of the ParA-SopA families can act as transcriptional repressors and are themselves often regulated by members of the ParB-SopB family (7,9,24,27,36).Spo0J is required both for accurate chromosome partitioning and for sporulation (14). Spo0J binds to a series of sites (parS sites) in the origin-proximal 20% of the chromosome (22), clustering them into a large focus (10,21,23). Defects in spo0J perturb chromosome partitioning. Cultures of spo0J null mutant cells produce approximately 1% anucleate cells, a frequency approximately 20-to 100-fold greater than that of wildtype cells (14).A null mutation in spo0J also causes an approximately 300-fold defect in sporulation relative to wild-type cells (14,28,34). spo0J null mutants ar...

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“…The exact functions of the other spoO genes remains unknown. However, the amino acid sequence homology of SpoOA and SpoOF to the transcriptionally related proteins of dual regulatory factors (18,30,34) suggests the possibility that SpoOA itself may have some role in regulating transcription.…”

Section: And Discussionmentioning

confidence: 99%

Mapping of sporulation-deficient suppressor mutations for a catabolite-resistant sporulation (crsA) mutant of the major sigma-43 factor of Bacillus subtilis.

Kawamura

Yamashita

Kobayashi

et al. 1989

J. Gen. Appl. Microbiol.

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Extragenic suppressor mutants for the sigA47 (crsA47, rpoD47) mutation were isolated and analyzed. Among 48 spontaneous suppressor mutants (sca),13 were deficient in sporulation and these mutations were mapped in the spo0A, spoOB, and spoOF genes, respectively. During the mapping studies, spoOG14 and spoOD8 mutations were found to be alleles of the spoOA and spoOB genes, respectively. These results indicate that a link exists between the RNA polymerase sigA enzyme, glucose catabolite repression, and some of the spoO gene functions.The sporulation of Bacillus subtilis is blocked at very early stages in the presence of relatively high concentrations of glucose (7,20,29). Takahashi (28) has isolated a number of mutants that overcome this repressive effect of glucose on sporulation. These mutations were located at six distinct loci, crsA to crsF, on the chromosome (25). Recently, it has been shown that three crsA mutations, crsAl, crsA4, and crsA47, are identical two base-changes, CCT (proline) to TTT (phenylalanine), within the coding sequence of the RNA polymerase 543 gene (rpoD or sigA) (8,12,17), and can suppress various sporulation defects (12, 15).

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Conserved domains in bacterial regulatory proteins that respond to environmental stimuli

Cited by 468 publications

References 0 publications

Identification of phosphate starvation-inducible genes in Escherichia coli K-12 by DNA sequence analysis of psi::lacZ(Mu d1) transcriptional fusions

Identification of phosphate starvation-inducible genes in Escherichia coli K-12 by DNA sequence analysis of psi::lacZ(Mu d1) transcriptional fusions

Control of Sporulation Gene Expression in Bacillus subtilis by the Chromosome Partitioning Proteins Soj (ParA) and Spo0J (ParB)

Mapping of sporulation-deficient suppressor mutations for a catabolite-resistant sporulation (crsA) mutant of the major sigma-43 factor of Bacillus subtilis.

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