Dataset Information

Reciprocal transcriptional and posttranscriptional growth-phase-dependent expression of sfh, a gene that encodes a paralogue of the nucleoid-associated protein H-NS.

ABSTRACT: The IncHI1 self-transmissible plasmid pSf-R27 from Shigella flexneri 2a strain 2457T harbors sfh, a gene that codes for a protein with strong amino acid sequence homology to the global transcription regulator and nucleoid-associated protein H-NS and to its paralogue, StpA. Previously, we discovered that the expression of sfh mRNA is growth phase dependent such that in cultures growing in Lennox broth at 37 degrees C, the transcript is readily detectable in the early stages of exponential growth but is not detectable at the onset of stationary phase. In contrast, the Sfh protein is poorly expressed in early-exponential growth when sfh mRNA is abundant whereas it is expressed to a high level in early stationary phase, when sfh transcript expression is low (P. Deighan, C. Beloin, and C. J. Dorman, Mol. Microbiol. 48:1401-1416, 2003). This unusual pattern of reciprocal mRNA and protein expression is not due to growth phase-dependent effects on either mRNA or protein stability, nor is it due to the known abilities of the Sfh, StpA, and H-NS proteins to influence sfh gene expression. Instead, our data point to a blockade of sfh mRNA translation in early-exponential growth that is relieved as the culture enters the stationary phase of growth. Replacing the 5' end and translation initiation signals of the sfh mRNA with heterologous sequences did not alter the growth phase-dependent expression of the Sfh protein, suggesting that growth phase control of translation is intrinsic to another component of the message.

SUBMITTER: Doyle M

PROVIDER: S-EPMC1636274 | biostudies-other | 2006 Nov

REPOSITORIES: biostudies-other

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Publications

Reciprocal transcriptional and posttranscriptional growth-phase-dependent expression of sfh, a gene that encodes a paralogue of the nucleoid-associated protein H-NS.

Doyle Marie M Dorman Charles J CJ

Journal of bacteriology 20060825 21

The IncHI1 self-transmissible plasmid pSf-R27 from Shigella flexneri 2a strain 2457T harbors sfh, a gene that codes for a protein with strong amino acid sequence homology to the global transcription regulator and nucleoid-associated protein H-NS and to its paralogue, StpA. Previously, we discovered that the expression of sfh mRNA is growth phase dependent such that in cultures growing in Lennox broth at 37 degrees C, the transcript is readily detectable in the early stages of exponential growth ...[more]

PMID: 16936022

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Project description:Transposon insertion sequencing (TIS; also known as TnSeq) is a potent approach commonly used to comprehensively define the genetic loci that contribute to bacterial fitness in diverse environments. A key presumption underlying analyses of TIS datasets is that loci with a low frequency of transposon insertions contribute to fitness. However, it is not known whether factors such as nucleoid binding proteins can alter the frequency of transposon insertion and thus whether TIS output may systematically reflect factors that are independent of the role of the loci in fitness. Here, we investigated whether the histone-like nucleoid structuring (H-NS) protein, which preferentially associates with AT-rich sequences, modulates the frequency of Mariner transposon insertion in the Vibrio cholerae genome, using comparative analysis of TIS results from wild-type (wt) and Δhns V. cholerae strains. These analyses were overlaid on gene classification based on GC content as well as on extant genome-wide identification of H-NS binding loci. Our analyses revealed a significant dearth of insertions within AT-rich loci in wt V. cholerae that was not apparent in the Δhns insertion library. Additionally, we observed a striking correlation between genetic loci that are overrepresented in the Δhns insertion library relative to their insertion frequency in wt V. cholerae and loci previously found to physically interact with H-NS. Collectively, our findings reveal that factors other than genetic fitness can systematically modulate the frequency of transposon insertions in TIS studies and add a cautionary note to interpretation of TIS data, particularly for AT-rich sequences. Transposon insertion sequencing (TIS) is often used to assess the relative frequency with which genetic loci can be disrupted, which is taken as an indicator of their importance for bacterial fitness. Here, we report that biological factors other than the relative levels of fitness of insertion mutants can influence TIS output. We found that the presence of the DNA binding protein H-NS, which preferentially recognizes AT-rich sequences, is linked to significant underrepresentation of mutations within AT-rich loci in transposon insertion libraries. Furthermore, there is a marked correspondence between loci bound by H-NS and loci with an increased frequency of disruption in a Δhns insertion library relative to a wt library. Our data suggest that factors other than genetic fitness (e.g., DNA binding proteins such as H-NS) can systematically modulate the frequency of transposon insertions in TIS studies and add a note of caution for interpretation of TIS data.

Dataset Information

Reciprocal transcriptional and posttranscriptional growth-phase-dependent expression of sfh, a gene that encodes a paralogue of the nucleoid-associated protein H-NS.

Publications

Reciprocal transcriptional and posttranscriptional growth-phase-dependent expression of sfh, a gene that encodes a paralogue of the nucleoid-associated protein H-NS.

Similar Datasets

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