Rny

• Description: RNase Y, 5' end sensitive endoribonuclease, involved in the degradation/processing of mRNA
  
  

Gene name	rny
Synonyms	ymdA
Essential	yes
Product	RNase Y
Function	RNA processing and degradation
Gene expression levels in SubtiExpress: rny
Interactions involving this protein in SubtInteract: Rny
Regulatory function of this protein in SubtiPathways: Central C-metabolism
MW, pI	58,7 kDa, 5.39
Gene length, protein length	1560 bp, 520 amino acids
Immediate neighbours	pbpX, ymdB
Sequences	Protein DNA DNA_with_flanks
Genetic context This image was kindly provided by SubtiList
Expression at a glance   PubMed

Categories containing this gene/protein

Rnases, biofilm formation, essential genes, membrane proteins

The gene

Basic information

• Locus tag: BSU16960

Phenotypes of a mutant

• essential PubMed
• transcription profile resulting from rny depletion: GEO PubMed
• defect in spore germination PubMed
• a study from the lab of Ciaran Condon reports that rny is non-essential and that the mutant is strongly impaired in sporulation, genetic competence and many other phenotypes PubMed

Database entries

• DBTBS entry: no entry

• SubtiList entry: [1]

• GEO entry: [2] (rny depletion vs. normal rny expression) PubMed

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity:
  • RNase Y cleaves S-box riboswitch RNAs in vivo and in vitro PubMed
  • preference for 5' monophosphorylated substrate in vitro PubMed
  • endonucleolytic cleavage PubMed
  • required for the processing of the gapA operon mRNA PubMed
  • cleavage activity appears sensitive to downstream secondary structure PubMed
  • RNase Y initiates the degradation of rpsO mRNA PubMed
  • RNase Y is responsible for the degradation of 23S rRNA, 16S rRNA, and mRNAs in aging spores PubMed

• Protein family: Member of the HD superfamily of metal-dependent phosphohydrolases; 2',3' cyclic nucleotide phosphodiesterase family (according to Swiss-Prot)

• Paralogous protein(s):

Extended information on the protein

• Kinetic information:

• Domains:
  • transmembrane domain (aa 5–24) PubMed
  • coiled-coiled domain (may form a leucine zipper) (aa 30-150) PubMed
  • KH domain (aa 210–280) PubMed
  • HD domain (aa 330–430) PubMed
  • C-terminal domain (aa 430-520) PubMed

• Modification:

• Cofactor(s): requires Mg+2, which can be replaced by Zn+2 or Mn+2 ions, PubMed

• Effectors of protein activity: appears sensitive to downstream secondary structure, PubMed

• Interactions:
  • RNase Y forms dimers PubMed
  • part of the RNA degradosome
  • RNase Y-PfkA PubMed
  • RNase Y-Eno PubMed, K(D) of the interaction: 100 nM PubMed
  • RNase Y-PnpA PubMed, K(D) of the interaction: 5 nM PubMed
  • RNase Y-RnjA PubMed
  • RNase Y-CshA PubMed
  • DynA-RNase Y PubMed

• Localization:
  • cell membrane, single-pass membrane protein PubMed
  • forms foci at the site of septation PubMed

Database entries

• Structure:

• UniProt: O31774

• KEGG entry: [3]

• E.C. number: 3.1.4.16

Additional information

required for the processing of the gapA operon mRNA

Expression and regulation

• Operon:
  • rny-ymdB PubMed
  • rny PubMed

• Expression browser: rny PubMed

• Sigma factor:

• Regulation: constitutive

• Regulatory mechanism:

• Additional information:
  • there is a terminator between rny and ymdB, most transcripts terminate there PubMed
  • The mRNA has a long 5' leader region. This may indicate RNA-based regulation PubMed

Biological materials

• Mutant:
  • essential!!!!
  • 4043 (rny under p-spac control, cat), GP193 (rny under p-xyl control, cat), both available in Jörg Stülke's lab
  • SSB447 (rny under P-spac control, "erm") available in Putzer lab.

• Expression vector:
  • N-terminal Strep-tag, expression in E. coli, in pGP172: pGP441, available in Jörg Stülke's lab
  • N-terminal Strep-tag, for SPINE, expression in B. subtilis, in pGP380: pGP775, available in Jörg Stülke's lab
  • C-terminal Strep-tag, for SPINE, expression in B. subtilis, in pGP382: pGP1852, available in Jörg Stülke's lab
  • Expression of RNase Y missing the N-terminal transmembrane domain (25aa) as an intein fusion in E. coli (no tag left in the purified protein) available in the Putzer lab
  • wild type rny, expression in B. subtilis, in pBQ200: pGP1201, available in Jörg Stülke's lab
  • there is also a series of domain constructs present in pBQ200, all available in Jörg Stülke's lab
  • chromosomal expression of Rny-Strep, spc: GP1033, available in Jörg Stülke's lab

• lacZ fusion: pGP459 (in pAC7), available in Jörg Stülke's lab

• GFP fusion:
  • B. subtilis 3569 (amyE:: (p-xyl rny-gfpmut1-spc)), available in Errington lab
  • pGP1368 for chromosomal expression of rny-YFP, available in Jörg Stülke's lab

• two-hybrid system: B. pertussis adenylate cyclase-based bacterial two hybrid system (BACTH), available in Jörg Stülke's lab

• FLAG-tag construct: GP1030 (spc, based on pGP1331), available in Jörg Stülke's lab

• Antibody: available in van Dijl and in Jörg Stülke's lab

Labs working on this gene/protein

Harald Putzer, IBPC Paris, France Homepage

Jörg Stülke, University of Göttingen, Germany Homepage