Dataset Information

Activity of Rhodobacter sphaeroides RpoHII, a second member of the heat shock sigma factor family.

ABSTRACT: We have identified a second RpoH homolog, RpoH(II), in the alpha-proteobacterium Rhodobacter sphaeroides. Primary amino acid sequence comparisons demonstrate that R. sphaeroides RpoH(II) belongs to a phylogenetically distinct group with RpoH orthologs from alpha-proteobacteria that contain two rpoH genes. Like its previously identified paralog, RpoH(I), RpoH(II) is able to complement the temperature-sensitive phenotype of an Escherichia coli sigma(32) (rpoH) mutant. In addition, we show that recombinant RpoH(I) and RpoH(II) each transcribe two E. coli sigma(32)-dependent promoters (rpoD P(HS) and dnaK P1) when reconstituted with E. coli core RNA polymerase. We observed differences, however, in the ability of each sigma factor to recognize six R. sphaeroides promoters (cycA P1, groESL(1), rpoD P(HS), dnaK P1, hslO, and ecfE), all of which resemble the E. coli sigma(32) promoter consensus. While RpoH(I) reconstituted with R. sphaeroides core RNA polymerase transcribed all six promoters, RpoH(II) produced detectable transcripts from only four promoters (cycA P1, groESL(1), hslO, and ecfE). These results, in combination with previous work demonstrating that an RpoH(I) mutant mounts a typical heat shock response, suggest that while RpoH(I) and RpoH(II) have redundant roles in response to heat, they may also have roles in response to other environmental stresses.

SUBMITTER: Green HA

PROVIDER: S-EPMC1540091 | biostudies-literature | 2006 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Activity of Rhodobacter sphaeroides RpoHII, a second member of the heat shock sigma factor family.

Green Heather A HA Donohue Timothy J TJ

Journal of bacteriology 20060801 16

We have identified a second RpoH homolog, RpoH(II), in the alpha-proteobacterium Rhodobacter sphaeroides. Primary amino acid sequence comparisons demonstrate that R. sphaeroides RpoH(II) belongs to a phylogenetically distinct group with RpoH orthologs from alpha-proteobacteria that contain two rpoH genes. Like its previously identified paralog, RpoH(I), RpoH(II) is able to complement the temperature-sensitive phenotype of an Escherichia coli sigma(32) (rpoH) mutant. In addition, we show that rec ...[more]

PMID: 16885439

Similar Datasets

Project description:We report the role of a gene (rpoH) from the facultative phototroph Rhodobacter sphaeroides that encodes a protein (sigma37) similar to Escherichia coli sigma32 and other members of the heat shock family of eubacterial sigma factors. R. sphaeroides sigma37 controls genes that function during environmental stress, since an R. sphaeroides deltaRpoH mutant is approximately 30-fold more sensitive to the toxic oxyanion tellurite than wild-type cells. However, the deltaRpoH mutant lacks several phenotypes characteristic of E. coli cells lacking sigma32. For example, an R. sphaeroides deltaRpoH mutant is not generally defective in phage morphogenesis, since it plates the lytic virus RS1, as well as its wild-type parent. In characterizing the response of R. sphaeroides to heat, we found that its growth temperature profile is different when cells generate energy by aerobic respiration, anaerobic respiration, or photosynthesis. However, growth of the deltaRpoH mutant is comparable to that of a wild-type strain under each of these conditions. The deltaRpoH mutant mounted a heat shock response when aerobically grown cells were shifted from 30 to 42 degrees C, but it exhibited altered induction kinetics of approximately 120-, 85-, 75-, and 65-kDa proteins. There was also reduced accumulation of several presumed heat shock transcripts (rpoD P(HS), groESL1, etc.) when aerobically grown deltaRpoH cells were placed at 42 degrees C. Under aerobic conditions, it appears that another sigma factor enables the deltaRpoH mutant to mount a heat shock response, since either RNA polymerase preparations from an deltaRpoH mutant, reconstituted Esigma37, or a holoenzyme containing a 38-kDa protein (sigma38) each transcribed E. coli Esigma32-dependent promoters. The lower growth temperature profile of photosynthetic cells is correlated with a difference in heat-inducible gene expression, since neither wild-type cells or the deltaRpoH mutant mount a typical heat shock response after such cultures were shifted from 30 to 37 degrees C.

Project description:Cells reprogram their transcriptome in response to stress, such as heat shock. In free-living bacteria, the transcriptomic reprogramming is mediated by increased DNA-binding activity of heat shock sigma factors and activation of genes normally repressed by heat-induced transcription factors. In this study, we performed transcriptomic analyses to investigate heat shock response in the obligate intracellular bacterium Chlamydia trachomatis, whose genome encodes only three sigma factors and a single heat-induced transcription factor. Nearly one-third of C. trachomatis genes showed statistically significant (≥1.5-fold) expression changes 30 min after shifting from 37 to 45°C. Notably, chromosomal genes encoding chaperones, energy metabolism enzymes, type III secretion proteins, as well as most plasmid-encoded genes, were differentially upregulated. In contrast, genes with functions in protein synthesis were disproportionately downregulated. These findings suggest that facilitating protein folding, increasing energy production, manipulating host activities, upregulating plasmid-encoded gene expression, and decreasing general protein synthesis helps facilitate C. trachomatis survival under stress. In addition to relieving negative regulation by the heat-inducible transcriptional repressor HrcA, heat shock upregulated the chlamydial primary sigma factor σ66 and an alternative sigma factor σ28. Interestingly, we show for the first time that heat shock downregulates the other alternative sigma factor σ54 in a bacterium. Downregulation of σ54 was accompanied by increased expression of the σ54 RNA polymerase activator AtoC, thus suggesting a unique regulatory mechanism for reestablishing normal expression of select σ54 target genes. Taken together, our findings reveal that C. trachomatis utilizes multiple novel survival strategies to cope with environmental stress and even to replicate. Future strategies that can specifically target and disrupt Chlamydia's heat shock response will likely be of therapeutic value.

Dataset Information

Activity of Rhodobacter sphaeroides RpoHII, a second member of the heat shock sigma factor family.

Publications

Activity of Rhodobacter sphaeroides RpoHII, a second member of the heat shock sigma factor family.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets