DisA

• Description: DNA integrity scanning protein,diadenylate cyclase, delays sporulation in the case of chromosome damage, the DisA-dependent checkpoint arrests DNA replication during B. subtilis spore outgrowth until the germinating spore's genome is free of damage
  
  

Gene name	disA
Synonyms	yacK
Essential	no
Product	DNA integrity scanning protein, has diadenylate cyclase activity
Function	control of sporulation initiation
Gene expression levels in SubtiExpress: disA
Interactions involving this protein in SubtInteract: DisA
Metabolic function and regulation of this protein in SubtiPathways: disA
MW, pI	40 kDa, 5.569
Gene length, protein length	1080 bp, 360 aa
Immediate neighbours	radA, yacL
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

cell division, metabolism of signalling nucleotides, sporulation/ other, general stress proteins (controlled by SigB), cell envelope stress proteins (controlled by SigM, V, W, X, Y), heat shock proteins

This gene is a member of the following regulons

CtsR regulon, SigB regulon, SigM regulon

The gene

Basic information

• Locus tag: BSU00880

Database entries

• BsubCyc: BSU00880

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity:
  • synthesis of c-di-AMP from two molecules of ATP PubMed

• Protein family: disA family (according to Swiss-Prot)

• Paralogous protein(s):

Extended information on the protein

• Kinetic information:

• Domains:
  • contains a DAC domain for the synthesis of c-di-AMP PubMed

• Modification:

• Cofactors:

• Effectors of protein activity:
  • RadA is an inhibitor of DisA activity PubMed

• Interactions:
  • forms octamers
  • RacA-DisA PubMed
  • RadA-DisA PubMed, this interaction result in the inhibition of DisA activity PubMed

• Localization:
  • see a video showing the movement of DisA in the cell (in real time) PubMed
  • forms discrete globular foci in germinating spres that colocalize with the nucleoid PubMed

Database entries

• BsubCyc: BSU00880

• Structure: 3C23 PubMed

• UniProt: P37573

• KEGG entry: [3]

• E.C. number:

Expression and regulation

• Operon:
  • ctsR-mcsA-mcsB-clpC-radA-disA PubMed
  • disA PubMed

• Expression browser: disA PubMed

• Sigma factor:
  • ctsR: SigA PubMed, SigB PubMed1 PubMed2
  • disA: SigM PubMed

• Regulation:
  • induced by stress (SigB) PubMed, induced by heat (CtsR) PubMed
  • expressed during sporulation in the mother cell PubMed
  • expressed during germination and spore outgrowth PubMed

• Regulatory mechanism: CtsR: transcription repression PubMed

• Additional information:
  • number of protein molecules per cell (minimal medium with glucose and ammonium): 314 PubMed
  • number of protein molecules per cell (complex medium with amino acids, without glucose): 393 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, exponential phase): 465 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, early stationary phase after glucose exhaustion): 179 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, late stationary phase after glucose exhaustion): 232 PubMed

Biological materials

• Mutant:
  • GP987 (disA::tet), available in Jörg Stülke's lab
  • BKG2 (radA-disA::spc), available in Jörg Stülke's lab
  • 1A939 ( disA::tet), PubMed, available at BGSC

• Expression vector:
  • IPTG inducible expression of His-disA in E. coli: pGP2563 (in pET19b), available in Jörg Stülke's lab

• lacZ fusion:

• GFP fusion:

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

• Antibody:

References

Reviews

Fabian M Commichau, Achim Dickmanns, Jan Gundlach, Ralf Ficner, Jörg Stülke
A jack of all trades: the multiple roles of the unique essential second messenger cyclic di-AMP.
Mol Microbiol: 2015, 97(2);189-204
[PubMed:25869574] [WorldCat.org] [DOI] (I p)

Justin S Lenhart, Jeremy W Schroeder, Brian W Walsh, Lyle A Simmons
DNA repair and genome maintenance in Bacillus subtilis.
Microbiol Mol Biol Rev: 2012, 76(3);530-64
[PubMed:22933559] [WorldCat.org] [DOI] (I p)

Ute Römling
Great times for small molecules: c-di-AMP, a second messenger candidate in Bacteria and Archaea.
Sci Signal: 2008, 1(33);pe39
[PubMed:18714086] [WorldCat.org] [DOI] (I e)

Original publications

Carolina Gándara, Juan C Alonso
DisA and c-di-AMP act at the intersection between DNA-damage response and stress homeostasis in exponentially growing Bacillus subtilis cells.
DNA Repair (Amst): 2015, 27;1-8
[PubMed:25616256] [WorldCat.org] [DOI] (I p)

Silvia S Campos, Juan R Ibarra-Rodriguez, Rocío C Barajas-Ornelas, Fernando H Ramírez-Guadiana, Armando Obregón-Herrera, Peter Setlow, Mario Pedraza-Reyes
Interaction of apurinic/apyrimidinic endonucleases Nfo and ExoA with the DNA integrity scanning protein DisA in the processing of oxidative DNA damage during Bacillus subtilis spore outgrowth.
J Bacteriol: 2014, 196(3);568-78
[PubMed:24244006] [WorldCat.org] [DOI] (I p)

Lei Zhang, Zheng-Guo He
Radiation-sensitive gene A (RadA) targets DisA, DNA integrity scanning protein A, to negatively affect cyclic Di-AMP synthesis activity in Mycobacterium smegmatis.
J Biol Chem: 2013, 288(31);22426-36
[PubMed:23760274] [WorldCat.org] [DOI] (I p)

Cao Zheng, Jieping Wang, Yunchao Luo, Yang Fu, Jianmei Su, Jin He
Highly efficient enzymatic preparation of c-di-AMP using the diadenylate cyclase DisA from Bacillus thuringiensis.
Enzyme Microb Technol: 2013, 52(6-7);319-24
[PubMed:23608499] [WorldCat.org] [DOI] (I p)

Felix M P Mehne, Katrin Gunka, Hinnerk Eilers, Christina Herzberg, Volkhard Kaever, Jörg Stülke
Cyclic di-AMP homeostasis in bacillus subtilis: both lack and high level accumulation of the nucleotide are detrimental for cell growth.
J Biol Chem: 2013, 288(3);2004-17
[PubMed:23192352] [WorldCat.org] [DOI] (I p)

Yaara Oppenheimer-Shaanan, Ezequiel Wexselblatt, Jehoshua Katzhendler, Eylon Yavin, Sigal Ben-Yehuda
c-di-AMP reports DNA integrity during sporulation in Bacillus subtilis.
EMBO Rep: 2011, 12(6);594-601
[PubMed:21566650] [WorldCat.org] [DOI] (I p)

Gregor Witte, Sophia Hartung, Katharina Büttner, Karl-Peter Hopfner
Structural biochemistry of a bacterial checkpoint protein reveals diadenylate cyclase activity regulated by DNA recombination intermediates.
Mol Cell: 2008, 30(2);167-78
[PubMed:18439896] [WorldCat.org] [DOI] (I p)

Adrian J Jervis, Penny D Thackray, Chris W Houston, Malcolm J Horsburgh, Anne Moir
SigM-responsive genes of Bacillus subtilis and their promoters.
J Bacteriol: 2007, 189(12);4534-8
[PubMed:17434969] [WorldCat.org] [DOI] (P p)

Michal Bejerano-Sagie, Yaara Oppenheimer-Shaanan, Idit Berlatzky, Alex Rouvinski, Mor Meyerovich, Sigal Ben-Yehuda
A checkpoint protein that scans the chromosome for damage at the start of sporulation in Bacillus subtilis.
Cell: 2006, 125(4);679-90
[PubMed:16713562] [WorldCat.org] [DOI] (P p)

Erik Boye
DisA, a busy bee that monitors chromosome integrity.
Cell: 2006, 125(4);641-3
[PubMed:16713555] [WorldCat.org] [DOI] (P p)

Sigal Ben-Yehuda, David Z Rudner, Richard Losick
RacA, a bacterial protein that anchors chromosomes to the cell poles.
Science: 2003, 299(5606);532-6
[PubMed:12493822] [WorldCat.org] [DOI] (I p)

A Petersohn, M Brigulla, S Haas, J D Hoheisel, U Völker, M Hecker
Global analysis of the general stress response of Bacillus subtilis.
J Bacteriol: 2001, 183(19);5617-31
[PubMed:11544224] [WorldCat.org] [DOI] (P p)

I Derré, G Rapoport, T Msadek
CtsR, a novel regulator of stress and heat shock response, controls clp and molecular chaperone gene expression in gram-positive bacteria.
Mol Microbiol: 1999, 31(1);117-31
[PubMed:9987115] [WorldCat.org] [DOI] (P p)

E Krüger, T Msadek, M Hecker
Alternate promoters direct stress-induced transcription of the Bacillus subtilis clpC operon.
Mol Microbiol: 1996, 20(4);713-23
[PubMed:8793870] [WorldCat.org] [DOI] (P p)