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A novel sigma factor reveals a unique regulon controlling cell-specific recombination in Mycoplasma genitalium.

Torres-Puig S, Broto A, Querol E, Piñol J, Pich OQ - Nucleic Acids Res. (2015)

Bottom Line: Herein, we found that MG428 activates transcription of recA, ruvA and ruvB as well as several genes with unknown function.Additionally, MG428 was found to interact with the RNAP core enzyme, reinforcing the predicted role of this protein as an alternative sigma factor.Finally, our results indicate that MG428 contributes to the generation of genetic diversity in this model organism.

View Article: PubMed Central - PubMed

Affiliation: Institut de Biotecnologia i Biomedicina and Departament de Bioquímica i Biologia Molecular. Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.

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Analysis of gene expression by qRT-PCR. Transcriptional analysis of selected M. genitalium genes in the WT and several mutant strains. Three independent biological repeats were performed and the respective fold-changes in gene expression are indicated with diamonds, squares and triangles. Mean fold-changes for each target gene are represented by color bars. Statistical significance of mean fold-changes above the cutoff (>2) was assessed with Student's t test. Statistically significant values (P < 0.05) are indicated with a red asterisk. Transcription of MG_220 (309 bp) in the Tn::MG_220 mutant could not be assessed by qRT-PCR due to the presence of a TnCatMG_428 MiniTnp insertion in this gene.
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Figure 3: Analysis of gene expression by qRT-PCR. Transcriptional analysis of selected M. genitalium genes in the WT and several mutant strains. Three independent biological repeats were performed and the respective fold-changes in gene expression are indicated with diamonds, squares and triangles. Mean fold-changes for each target gene are represented by color bars. Statistical significance of mean fold-changes above the cutoff (>2) was assessed with Student's t test. Statistically significant values (P < 0.05) are indicated with a red asterisk. Transcription of MG_220 (309 bp) in the Tn::MG_220 mutant could not be assessed by qRT-PCR due to the presence of a TnCatMG_428 MiniTnp insertion in this gene.

Mentions: The existence of transcriptional changes between the WT strain and the different MG_428 mutants described above was analyzed by qRT-PCR (Figure 3). As expected, no MG_428 transcript could be detected in the ΔMG_428 mutant. However, in agreement with the western blot analysis, we found increased levels of MG_428 transcript (∼20-fold) in the complemented mutants as compared to the WT strain. No transcriptional changes of the MG_427 gene were observed in any of the strains tested, suggesting that MG428 overexpression has no effect on MG_427 transcript levels.


A novel sigma factor reveals a unique regulon controlling cell-specific recombination in Mycoplasma genitalium.

Torres-Puig S, Broto A, Querol E, Piñol J, Pich OQ - Nucleic Acids Res. (2015)

Analysis of gene expression by qRT-PCR. Transcriptional analysis of selected M. genitalium genes in the WT and several mutant strains. Three independent biological repeats were performed and the respective fold-changes in gene expression are indicated with diamonds, squares and triangles. Mean fold-changes for each target gene are represented by color bars. Statistical significance of mean fold-changes above the cutoff (>2) was assessed with Student's t test. Statistically significant values (P < 0.05) are indicated with a red asterisk. Transcription of MG_220 (309 bp) in the Tn::MG_220 mutant could not be assessed by qRT-PCR due to the presence of a TnCatMG_428 MiniTnp insertion in this gene.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4446450&req=5

Figure 3: Analysis of gene expression by qRT-PCR. Transcriptional analysis of selected M. genitalium genes in the WT and several mutant strains. Three independent biological repeats were performed and the respective fold-changes in gene expression are indicated with diamonds, squares and triangles. Mean fold-changes for each target gene are represented by color bars. Statistical significance of mean fold-changes above the cutoff (>2) was assessed with Student's t test. Statistically significant values (P < 0.05) are indicated with a red asterisk. Transcription of MG_220 (309 bp) in the Tn::MG_220 mutant could not be assessed by qRT-PCR due to the presence of a TnCatMG_428 MiniTnp insertion in this gene.
Mentions: The existence of transcriptional changes between the WT strain and the different MG_428 mutants described above was analyzed by qRT-PCR (Figure 3). As expected, no MG_428 transcript could be detected in the ΔMG_428 mutant. However, in agreement with the western blot analysis, we found increased levels of MG_428 transcript (∼20-fold) in the complemented mutants as compared to the WT strain. No transcriptional changes of the MG_427 gene were observed in any of the strains tested, suggesting that MG428 overexpression has no effect on MG_427 transcript levels.

Bottom Line: Herein, we found that MG428 activates transcription of recA, ruvA and ruvB as well as several genes with unknown function.Additionally, MG428 was found to interact with the RNAP core enzyme, reinforcing the predicted role of this protein as an alternative sigma factor.Finally, our results indicate that MG428 contributes to the generation of genetic diversity in this model organism.

View Article: PubMed Central - PubMed

Affiliation: Institut de Biotecnologia i Biomedicina and Departament de Bioquímica i Biologia Molecular. Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain.

Show MeSH
Related in: MedlinePlus