rnhC

168

RNase HIII, endoribonuclease

Locus: BSU_28620

Molecular weight: 33.92 kDa

Isoelectric point: 10.07

Protein length: 313 aa

Gene length: 939 bp

Function: endonucleolytic cleavage of RNA in RNA-DNA hybrid molecules, processing of R-loops, maturation of Okazaki fragments

Product: Mn²⁺-dependent RNase HIII

Essential: no

E.C.: 3.1.26.4

Synonyms: rnhC, ysgB

Genomic Context

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List of homologs in different organisms, belongs to COG1039 (Galperin et al., 2021)

Gene

Coordinates: 2,926,031 → 2,926,972

Phenotypes of a mutant

• cold-sensisitive due to a defect in Okazaki fragment maturation PubMed
• a recA rnhC double mutant is not viable PubMed
• a rnhC polA double mutant is not viable at 25°C PubMed
• a rnhC fenA double mutant is not viable at 25°C PubMed
• sensitive to hydroxyurea PubMed
• a ''rnhB rnhC'' double mutant grows poorly PubMed
• defective in Swarming motility (prolonged period of immotility prior to Swarming) PubMed

The protein

Catalyzed reaction/ biological activity

• maturation of Okazaki fragments (removal of RNA) (together with DNA polymerase I) PubMed
• Endonucleolytic cleavage to 5'-phosphomonoester (according to UniProt)

Protein family

• RNase HII family (with RnhB, according to UniProt)

Cofactors

• Mn²⁺ PubMed

Structure

• 2D0B (PDB) (complex with Mg²⁺, Geobacillus stearothermophilus , 47% identity) PubMed
• 2D0A (PDB) (Geobacillus stearothermophilus, 47% identity) PubMed
• P94541 (AlphaFold)

Localization

• recruited to the replication forks PubMed

Expression and Regulation

Operons

• Genes: rnhC

  Description: PubMed

Biological materials

Mutant

• MGNA-B002 (ysgB::erm), available at the NBRP B. subtilis, Japan
• BP430 (ΔrnhC::cat) available in Fabian Commichau's lab
• BKE28620 (ΔrnhC::erm  trpC2) available at BGSC,  PubMed, upstream reverse: _UP1_CACGTAATAATCTCCTTTTT,  downstream forward: _UP4_TAGAAAAAAGCTTGCAGATT
• BKK28620 (ΔrnhC::kan  trpC2) available at BGSC,  PubMed, upstream reverse: _UP1_CACGTAATAATCTCCTTTTT,  downstream forward: _UP4_TAGAAAAAAGCTTGCAGATT

Expression vectors

• for expression/ purification from B. subtilis with C-terminal Strep-tag, for SPINE, expression from plasmid: pBP502 (E. coli amp & B. subtilis E/L) , available in Fabian Commichau's lab

References

Reviews

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Original Publications

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