rnhB

RNase HII, endoribonuclease, responsible for removal of single rNMPs incorporated into DNA by DNA polymerase during DNA replication

locus: BSU_16060

pI: 5.52

mw: 28.20 kDa

protein length: 255 aa

gene length: 768 bp

function: endonucleolytic cleavage of RNA in RNA-DNA hybrid molecules

product: RNase HII

essential: no

ec: 3.1.26.4

synonyms: rnh

Genomic Context

categories

• [category|SW 3|Information processing] → [category|SW 3.1|Genetics] → [category|SW 3.1.1|DNA replication]
• [category|SW 3|Information processing] → [category|SW 3.2|RNA synthesis and degradation] → [category|SW 3.2.4|RNases] → [category|SW 3.2.4.2|Endoribonucleases]

Gene

Coordinates: 1,677,451 → 1,678,218

Phenotypes of a mutant

• increased mutation rate [Pubmed|23882084], strand- and sequence-context–dependent GC → AT transitions [pubmed|29078353]
• a ''[gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB] [gene|B8B0F6B129475FDEEA8D538A51C153BF9A35887D|rnhC]'' double mutant grows poorly [Pubmed|23882084]
• a ''[gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB] [gene|E2A8B04CD729332F9E6A13195181983F751F2F25|mutS]-[gene|A86A013EDCE59D7EF84FB836A751E66C58CADF45|mutL]'' mutant is not viable [Pubmed|23882084]

The protein

Catalyzed reaction/ biological activity

• Endonucleolytic cleavage to 5'-phosphomonoester (according to UniProt)
• incises DNA-RNA duplexes at aingle rNMPs [pubmed|29078353]
• removal of single rNMPs incorporated into DNA by DNA polymerase during [SW|DNA replication] [Pubmed|23882084]

Protein family

• [SW|RNase] HII family (with [protein|B8B0F6B129475FDEEA8D538A51C153BF9A35887D|RnhC], according to UniProt)

[SW|Cofactors]

• Mg²⁺, Mn²⁺ [pubmed|29084857]

Structure

• [PDB|2ETJ] (from Thermotoga maritima, corresponding to aa 69 ... 250, 47% identity)

[SW|Localization]

• cytoplasm (according to Swiss-Prot)

Expression and Regulation

Operons

• genes: [gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB]-[gene|0D4A4A1B2FD9D0698D0A65010A8ABC84DA7386A2|ylqG]-[gene|3F271D9FFE4F30D845319CD71662695AA592EC8F|ylqH]
  description: [Pubmed|26577401]

  sigma factors

  • [protein|7024E4162A6D827069F882FDEACA696EBC05DD40|SigD]: sigma factor, [Pubmed|26577401], in [regulon|7024E4162A6D827069F882FDEACA696EBC05DD40|SigD regulon]

• genes: [gene|54CD247926F7FABE4ACE85EB7021B62500B47260|rbgA]-[gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB]-[gene|0D4A4A1B2FD9D0698D0A65010A8ABC84DA7386A2|ylqG]-[gene|3F271D9FFE4F30D845319CD71662695AA592EC8F|ylqH]
  description: [Pubmed|22383849]

Biological materials

Mutant

• BP423 (''ΔrnhB::aphA3'') available in [SW|Fabian Commichau]'s lab
• BKE16060 (Δ[gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB]::erm trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKE16060 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CACTTTCTCTTCTCCCTTAC, downstream forward: _UP4_TAAATCACCATGGACAAGGA
• BKK16060 (Δ[gene|D5275ECDC2A70FBAC2CA5980F04B272C3328FC90|rnhB]::kan trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKK16060 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CACTTTCTCTTCTCCCTTAC, downstream forward: _UP4_TAAATCACCATGGACAAGGA

Expression vectors

• for expression/ purification from ''B. subtilis'' with N-terminal Strep-tag, for [SW|SPINE], based on [SW|pGP380], expression from plasmid: pBP500 (E. coli amp & B. subtilis E/L) , available in [SW|Fabian Commichau]'s lab

two-hybrid system

• B. pertussis adenylate cyclase-based bacterial two hybrid system ([SW|BACTH]), available in [SW|Fabian Commichau]'s lab

References

Reviews

• 19228197,24464998,31464530

Original publications

• 9888800,10094689,12417313,17905985,23882084,26577401,29078353,29084857