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
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
Get the DNA and protein sequences (Barbe et al., 2009)
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

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
  • Rny-PfkA PubMed
  • Rny-Eno PubMed, K(D) of the interaction: 100 nM PubMed
  • Rny-PnpA PubMed, K(D) of the interaction: 5 nM PubMed
  • Rny-RnjA PubMed
  • Rny-CshA PubMed

• Localization:
  • cell membrane, single-pass membrane protein 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 Stülke 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 Stülke lab
  • N-terminal Strep-tag, for SPINE, expression in B. subtilis, in pGP380: pGP775 , available in Stülke 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 Stülke lab
  • there is also a series of domain constructs present in pBQ200, all available in Stülke lab
  • chromosomal expression of Rny-Strep, spc: GP1033, available in Jörg Stülke's lab

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

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

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

• Antibody: available in van Dijl and Stülke labs

Labs working on this gene/protein

Harald Putzer, IBPC Paris, France Homepage

Jörg Stülke, University of Göttingen, Germany Homepage

References

Reviews

Additional reviews: PubMed

Publications on B. subtilis rny

Additional publications: PubMed

Sylvain Durand, Laetitia Gilet, Philippe Bessières, Pierre Nicolas, Ciarán Condon
Three essential ribonucleases-RNase Y, J1, and III-control the abundance of a majority of Bacillus subtilis mRNAs.
PLoS Genet: 2012, 8(3);e1002520
[PubMed:22412379] [WorldCat.org] [DOI] (I p)

Einat Segev, Yoav Smith, Sigal Ben-Yehuda
RNA dynamics in aging bacterial spores.
Cell: 2012, 148(1-2);139-49
[PubMed:22209493] [WorldCat.org] [DOI] (I p)

Joseph A Newman, Lorraine Hewitt, Cecilia Rodrigues, Alexandra S Solovyova, Colin R Harwood, Richard J Lewis
Dissection of the network of interactions that links RNA processing with glycolysis in the Bacillus subtilis degradosome.
J Mol Biol: 2012, 416(1);121-36
[PubMed:22198292] [WorldCat.org] [DOI] (I p)

Gintaras Deikus, David H Bechhofer
5' End-independent RNase J1 endonuclease cleavage of Bacillus subtilis model RNA.
J Biol Chem: 2011, 286(40);34932-40
[PubMed:21862575] [WorldCat.org] [DOI] (I p)

Diethmaier C, Pietack N, Gunka K, Wrede C, Lehnik-Habrink M, Herzberg C, Hübner S, Stülke J    
A Novel Factor Controlling Bistability in Bacillus subtilis: The YmdB Protein Affects
Flagellin Expression and Biofilm Formation. 
J Bacteriol.: 2011, 193(21):5997-6007. 
PubMed:21856853

Lehnik-Habrink M, Schaffer M, Mäder U, Diethmaier C, Herzberg C, Stülke J  
RNA processing in Bacillus subtilis: identification of targets of the essential RNase Y. 
Mol Microbiol. 2011 81(6): 1459-1473. 
PubMed:21815947