cshA [2018-10-21 16:00:49]

DEAD-box RNA helicase, important for adaptation to low temperatures

locus: BSU04580

pI: 9.89

mw: 57.11 kDa

protein length: 511 aa

gene length: 1533 bp

function: RNA helicase

product: DEAD-box RNA helicase

essential: no

synonyms: ydbR

Genomic Context

categories

• [category|SW 3|Information processing] → [category|SW 3.2|RNA synthesis and degradation] → [category|SW 3.2.3|DEAD-box RNA helicases]
• [category|SW 3|Information processing] → [category|SW 3.3|Protein synthesis, modification and degradation] → [category|SW 3.3.1|Translation] → [category|SW 3.3.1.6|Ribosome assembly]
• [category|SW 4|Lifestyles] → [category|SW 4.3|Coping with stress] → [category|SW 4.3.5|Cold stress proteins]
• [category|SW 6|Groups of genes] → [category|SW 6.2|Membrane proteins]

Gene

Coordinates: 511,106 → 512,641

Phenotypes of a mutant

• poor growth at low temperatures (16 to 20°C) [Pubmed|23175651]
• reduced number of [SW|ribosome]s [Pubmed|23175651]
• no expression of the ''[gene|C87E74FB67B53DD1C68916AA121388C60CE66E92|frlB]-[gene|9E3FF9D0B33D7EFEA3B61FF863BB46CC6DFD4F9B|frlO]-[gene|8256CAAEC591E40A8CB172FF053E03BFFD5A4B54|frlN]-[gene|1F245030D0C81DC96FC0642199CA00579CF46D06|frlM]-[gene|C3823AFED3E5C3D1EA04C38D861239468C9E6349|frlD]'' operon [Pubmed|23175651]
• strongly increased expression of the ''[gene|6F21B5E9EEF67E96808A1DEFA3A80DA2A0FF9462|pftA]-[gene|F6C3B267106E7510FABECF0B13ABF222901689A9|pftB]'' operon [Pubmed|23175651]
• transcription profile resulting from ''[gene|872CCB5A49C9000BD95E4B0472556D5F60F7D7A4|rny]'' depletion: [http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE36877 GEO] [Pubmed|23175651]

The protein

Catalyzed reaction/ biological activity

• RNA helicase
• required for [SW|ribosome] assembly (biogenesis of the large subunit) [Pubmed|23175651]
• unwind duplex RNA with or without overhangs [pubmed|28238534]

Protein family

• helicase C-terminal domain (according to Swiss-Prot) [SW|DEAD-box RNA helicases]

Paralogous protein(s)

• [protein|EACD0C84AA3CE41EF05EAA3D25AA1153293EB562|DeaD], [protein|0362D937D985846AEE4D0DEA57B62F915E8D35E5|YfmL], [protein|C83BD80DE0FD187D07B914F64696438ED5130107|CshB]

Structure

• [PDB|5IVL] [Pubmed|28238534]

[SW|Localization]

• cytoplasma, colocalizes with the ribosomes [Pubmed|16352840,27708634], cell membrane [Pubmed|20572937]

Expression and Regulation

Operons

• genes: [gene|E169F9711635457EA65FA71CCE9D7FF29DD8DF02|cshA]
  description: [pubmed|22383849]

Biological materials

Mutant

• GP1035 (Δ[gene|E169F9711635457EA65FA71CCE9D7FF29DD8DF02|cshA]::aphA3), available in [SW|Jörg Stülke]'s lab [pubmed|23175651]
• GP1083 (Δ[gene|E169F9711635457EA65FA71CCE9D7FF29DD8DF02|cshA]::cat), available in [SW|Jörg Stülke]'s lab
• BKE04580 (Δ[gene|E169F9711635457EA65FA71CCE9D7FF29DD8DF02|cshA]::erm trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKE04580 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CACTTATAAAACTGCCCTTT, downstream forward: _UP4_TAATTTGATCGATTCAGAGC
• BKK04580 (Δ[gene|E169F9711635457EA65FA71CCE9D7FF29DD8DF02|cshA]::kan trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKK04580 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CACTTATAAAACTGCCCTTT, downstream forward: _UP4_TAATTTGATCGATTCAGAGC

Expression vector

• for expression/ purification from ''B. subtilis'' with C-terminal Strep-tag, for [SW|SPINE], in [SW|pGP382]: pGP1387, available in [SW|Jörg Stülke]'s lab
• for expression/ purification from ''B. subtilis'' with C-terminal Strep-tag, for [SW|SPINE], expression from the native chromomsomal site: GP1026 (aphA3), available in [SW|Jörg Stülke]'s lab
• for expression/ purification from ''E. coli'' with N-terminal His-tag, in [SW|pWH844]: pGP1386, available in [SW|Jörg Stülke]'s lab

GFP fusion

• pGP1369 for chromosomal expression of CshA-YFP, available in [SW|Jörg Stülke]'s lab
• ''B. subtilis'' GP1081 cshA-gfp spc, available in [SW|Jörg Stülke]'s lab,
• GP1721 (in [SW|pBP43]), expression of '' cshA-GFP''::''spc'' under the native promoter, available in [SW|Jörg Stülke]'s lab [Pubmed|27708634]

two-hybrid system

• B. pertussis adenylate cyclase-based bacterial two hybrid system ([SW|BACTH]), available in [SW|Jörg Stülke]'s lab, [pubmed|20572937]

FLAG-tag construct

• GP1010 (spc, based on [SW|pGP1331]), available in [SW|Jörg Stülke]'s lab
• GP1074 (tet), available in [SW|Jörg Stülke]'s lab

Labs working on this gene/protein

• [SW|Mohamed Marahiel], Marburg University, Germany [http://www.uni-marburg.de/fb15/fachgebiete/bio/marahiel?language_sync=1 homepage]

References

Reviews

• 22568516,29314657,19747077,25907111

Original publications

• 20572937,23175651,21803996,16352840,27708634,27965645,28238534