DNA sequencing coverage in wild type and RNase H deficient Bacillus subtilis cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Bacillus subtilis subsp. subtilis NCIB 3610 = ATCC 6051 = DSM 10
PROVIDER: GSE154586 | GEO | 2020/07/16
REPOSITORIES: GEO
Similar Datasets
Project description:RNA-DNA hybrids form throughout the chromosome during normal cell growth and under stress conditions. When left unresolved, RNA-DNA hybrids can slow replication fork progression, cause DNA breaks, increase mutagenesis, and reduce gene expression. To remove hybrids, all organisms use ribonuclease H (RNase H) to specifically degrade the RNA portion of RNA-DNA hybrids. Here we show that, in addition to chromosomally encoded RNase HII and RNase HIII, Bacillus subtilis NCIB 3610 encodes a previously uncharacterized RNase HI protein, RnhP, on the endogenous plasmid pBS32. Like other RNase HI enzymes, RnhP incises Okazaki fragments, ribopatches, and a complementary RNA-DNA hybrid. We show that while chromosomally encoded RNase HIII is required for pBS32 hyper-replication, RnhP compensates for loss of RNase HIII activity on the chromosome. Consequently, loss of RnhP and RNase HIII impairs bacterial growth. We show that the decreased growth rate can be explained by laggard replication fork progression near the terminus region of the right replichore resulting in SOS-dependent inhibition of cell division. We conclude that B. subtilis NCIB 3610 encodes functional RNase HI, HII, and HIII, and pBS32-encoded RNase HI contributes to replication fork progression and chromosome stability while RNase HIII is important for chromosome stability and plasmid hyper-replication.
2020-07-16 | GSE154585 | GEO
Project description:RNA-DNA hybrids form throughout the chromosome during normal cell growth and under stress conditions. When left unresolved, RNA-DNA hybrids can slow replication fork progression, cause DNA breaks, increase mutagenesis, and reduce gene expression. To remove hybrids, all organisms use ribonuclease H (RNase H) to specifically degrade the RNA portion of RNA-DNA hybrids. Here we show that, in addition to chromosomally encoded RNase HII and RNase HIII, Bacillus subtilis NCIB 3610 encodes a previously uncharacterized RNase HI protein, RnhP, on the endogenous plasmid pBS32. Like other RNase HI enzymes, RnhP incises Okazaki fragments, ribopatches, and a complementary RNA-DNA hybrid. We show that while chromosomally encoded RNase HIII is required for pBS32 hyper-replication, RnhP compensates for loss of RNase HIII activity on the chromosome. Consequently, loss of RnhP and RNase HIII impairs bacterial growth. We show that the decreased growth rate can be explained by laggard replication fork progression near the terminus region of the right replichore resulting in SOS-dependent inhibition of cell division. We conclude that B. subtilis NCIB 3610 encodes functional RNase HI, HII, and HIII, and pBS32-encoded RNase HI contributes to replication fork progression and chromosome stability while RNase HIII is important for chromosome stability and plasmid hyper-replication.
2020-04-30 | GSE149552 | GEO
Project description:DNA sequencing coverage in wild type and RNase H deficient Bacillus subtilis cells
| PRJNA646729 | ENA
Project description:DNA sequencing coverage in wild type and RNase H deficient Bacillus subtilis cells [1]
| PRJNA629098 | ENA
Project description:DNA sequencing coverage in wild type and RNase H deficient Bacillus subtilis cells [2]
| PRJNA646731 | ENA
Project description:RNase J1 is the first nuclease with 5-3 exonuclease activity in bacteria and plays an important role in the maturation and degradation of mRNA. Absence of RNase J1 in cells has effect on cell morphology and growth rate. We did RNA-seq of Bacillus subtilis wild type and RNase J1 null strains (triplicates) to obtain more information about cellular role of this nuclease in cells. Complmentary ChIP-seq data have also been deposited in ArrayExpress under accession number E-MTAB-5659 ( https://www.ebi.ac.uk/arrayexpress/experiments/E-MTAB-5659 ).
2019-12-31 | E-MTAB-5660 | biostudies-arrayexpress
Project description:RNase Y and RNase E are disparate endoribonucleases that govern global mRNA turnover/processing in the two evolutionary distant bacteria Bacillus subtilis and Escherichia coli, respectively. The two enzymes share a similar in vitro cleavage specificity and subcellular localization. To evaluate the potential equivalence in biological function between the two enzymes in vivo we analyzed whether and to what extent RNase E is able to replace RNase Y in B. subtilis. Full-length RNase E almost completely restores wild type growth of the rny mutant. This is matched by a surprising reversal of transcript profiles both of individual genes and on a genome-wide scale. The single most important parameter to efficient complementation is the requirement for RNase E to localize to the inner membrane while truncation of the C-terminal sequences corresponding to the degradosome scaffold has only a minor effect. We also compared the in vitro cleavage activity for the major decay initiating ribonucleases Y, E and J and show that no conclusions can be drawn with respect to their activity in vivo. Our data confirm the notion that RNase Y and RNase E have evolved through convergent evolution towards a low specificity endonuclease activity universally important in bacteria.
| S-EPMC8096251 | biostudies-literature
Project description:This project is about the proteomes of wild type B. subtilis and its codY-null strain (The organism at right side was mislabeled as "E. coli")
2025-11-10 | PXD052459 | Pride
Project description:This project is about the proteomes of wild type B. subtilis and its codY-null strain (The organism at right side was mislabeled as "E. coli")
2025-11-10 | PXD055309 | Pride
Project description:The instability of messenger RNA is crucial to the control of gene expression. In Bacillus subtilis, RNase Y is the major decay-initiating endoribonuclease. Here, we show how this key enzyme regulates its own synthesis by modulating the longevity of its mRNA. Autoregulation is achieved through cleavages in two regions of the rny (RNase Y) transcript: (i) within the first ~100 nucleotides of the open reading frame, immediately inactivating the mRNA for further rounds of translation; (ii) cleavages in the rny 5' UTR, primarily within the 5'-terminal 50 nucleotides, creating entry sites for the 5' exonuclease J1 whose progression is blocked around position -15 of the rny mRNA, potentially by initiating ribosomes. This links the functional inactivation of the transcript by RNase J1 to translation efficiency, depending on the ribosome occupancy at the translation initiation site. By these mechanisms, RNase Y can initiate degradation of its own mRNA when the enzyme is not occupied with degradation of other RNAs and thus prevent its overexpression beyond the needs of RNA metabolism.
| S-EPMC10303841 | biostudies-literature