Hag

• Description: flagellin protein, about 20,000 subunits make up one flagellum
  
  

Gene name	hag
Synonyms
Essential	no
Product	flagellin protein
Function	motility and chemotaxis
Gene expression levels in SubtiExpress: hag
Interactions involving this protein in SubtInteract: Hag
Metabolic function and regulation of this protein in SubtiPathways: Hag
MW, pI	32 kDa, 4.782
Gene length, protein length	912 bp, 304 aa
Immediate neighbours	yvyC, csrA
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

motility and chemotaxis, most abundant proteins

This gene is a member of the following regulons

CodY regulon, CsrA regulon, ScoC regulon, SigD regulon

The gene

Basic information

• Locus tag: BSU35360

Phenotypes of a mutant

No swarming motility on B medium. PubMed

Database entries

• BsubCyc: BSU35360

• DBTBS entry: [1]

• SubtiList entry: [2]

The protein

Basic information/ Evolution

• Catalyzed reaction/ biological activity:

• Protein family: bacterial flagellin family (according to Swiss-Prot)

• Paralogous protein(s): YvzB (C-terminal domain of Hag)

Extended information on the protein

• Kinetic information:

• Domains:

• Modification:

• Cofactors:

• Effectors of protein activity:

• Interactions:
  • FlhA-Hag PubMed
  • FlhA-(FliS-Hag) PubMed
  • FliS-Hag PubMed
  • FliW-Hag PubMed

• Localization:
  • extracellular (no signal peptide), major component of the secretome PubMed
  • membrane PubMed

Database entries

• BsubCyc: BSU35360

• Structure: 3A5X (from Salmonella typhimurium, 42% identity)

• UniProt: P02968

• KEGG entry: [3]

• E.C. number:

Expression and regulation

• Operon: hag PubMed

• Expression browser: hag PubMed

• Sigma factor: SigD PubMed

• Regulation:
  • repressed during growth in the presence of branched chain amino acids (CodY) PubMed

• Regulatory mechanism:
  • CodY: transcription repression PubMed
  • CsrA: translation inhibition PubMed
  • ScoC: transcription repression PubMed

• Additional information:
  • the hag gene is strongly overexpressed in a ymdB mutant (loss of bistable gene expression) PubMed
  • belongs to the 100 most abundant proteins PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium): 150241 PubMed
  • number of protein molecules per cell (complex medium with amino acids, without glucose): 280953 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, exponential phase): 9552 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, early stationary phase after glucose exhaustion): 11147 PubMed
  • number of protein molecules per cell (minimal medium with glucose and ammonium, late stationary phase after glucose exhaustion): 24114 PubMed

Biological materials

• Mutant: GP901 (aphA3), GP902 (tet) PubMed, both available in Jörg Stülke's lab
  • 1A915 ( hag::cat), PubMed, available at BGSC
  • 1A783 ( hag::erm), PubMed, available at BGSC
  • 1A842 ( hag::kan), PubMed, available at BGSC

• Expression vector:
  • expression in B. subtilis, in pBQ200: pGP1089, available in Jörg Stülke's lab

• lacZ fusion: pGP1035 (in pAC6), pGP755 (in pAC7), there is also a series of promoter deletion variants in pAC6 and pAC7 PubMed, all available in Jörg Stülke's lab

• GFP fusion: BP494 (bglS:: (hag-promoter-cfp-aphA3)), BP496 (amyE:: (hag-promoter-iyfp-cat)), available in Jörg Stülke's lab

• two-hybrid system:

• Antibody:

References

Original Publications

Sampriti Mukherjee, Paul Babitzke, Daniel B Kearns
FliW and FliS function independently to control cytoplasmic flagellin levels in Bacillus subtilis.
J Bacteriol: 2013, 195(2);297-306
[PubMed:23144244] [WorldCat.org] [DOI] (I p)

Sampriti Mukherjee, Helen Yakhnin, Dave Kysela, Josh Sokoloski, Paul Babitzke, Daniel B Kearns
CsrA-FliW interaction governs flagellin homeostasis and a checkpoint on flagellar morphogenesis in Bacillus subtilis.
Mol Microbiol: 2011, 82(2);447-61
[PubMed:21895793] [WorldCat.org] [DOI] (I p)

Christine Diethmaier, Nico Pietack, Katrin Gunka, Christoph Wrede, Martin Lehnik-Habrink, Christina Herzberg, Sebastian Hübner, Jörg Stülke
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] [WorldCat.org] [DOI] (I p)

Kassem Hamze, Sabine Autret, Krzysztof Hinc, Soumaya Laalami, Daria Julkowska, Romain Briandet, Margareth Renault, Cédric Absalon, I Barry Holland, Harald Putzer, Simone J Séror
Single-cell analysis in situ in a Bacillus subtilis swarming community identifies distinct spatially separated subpopulations differentially expressing hag (flagellin), including specialized swarmers.
Microbiology (Reading): 2011, 157(Pt 9);2456-2469
[PubMed:21602220] [WorldCat.org] [DOI] (I p)

Gert Bange, Nico Kümmerer, Christoph Engel, Gunes Bozkurt, Klemens Wild, Irmgard Sinning
FlhA provides the adaptor for coordinated delivery of late flagella building blocks to the type III secretion system.
Proc Natl Acad Sci U S A: 2010, 107(25);11295-300
[PubMed:20534509] [WorldCat.org] [DOI] (I p)

Kassem Hamze, Daria Julkowska, Sabine Autret, Krzysztof Hinc, Krzysztofa Nagorska, Agnieszka Sekowska, I Barry Holland, Simone J Séror
Identification of genes required for different stages of dendritic swarming in Bacillus subtilis, with a novel role for phrC.
Microbiology (Reading): 2009, 155(Pt 2);398-412
[PubMed:19202088] [WorldCat.org] [DOI] (P p)

Prashant Kodgire, K Krishnamurthy Rao
hag expression in Bacillus subtilis is both negatively and positively regulated by ScoC.
Microbiology (Reading): 2009, 155(Pt 1);142-149
[PubMed:19118355] [WorldCat.org] [DOI] (P p)

Hannes Hahne, Susanne Wolff, Michael Hecker, Dörte Becher
From complementarity to comprehensiveness--targeting the membrane proteome of growing Bacillus subtilis by divergent approaches.
Proteomics: 2008, 8(19);4123-36
[PubMed:18763711] [WorldCat.org] [DOI] (I p)

Helen Yakhnin, Pallavi Pandit, Tom J Petty, Carol S Baker, Tony Romeo, Paul Babitzke
CsrA of Bacillus subtilis regulates translation initiation of the gene encoding the flagellin protein (hag) by blocking ribosome binding.
Mol Microbiol: 2007, 64(6);1605-20
[PubMed:17555441] [WorldCat.org] [DOI] (P p)

Christine Eymann, Annette Dreisbach, Dirk Albrecht, Jörg Bernhardt, Dörte Becher, Sandy Gentner, Le Thi Tam, Knut Büttner, Gerrit Buurman, Christian Scharf, Simone Venz, Uwe Völker, Michael Hecker
A comprehensive proteome map of growing Bacillus subtilis cells.
Proteomics: 2004, 4(10);2849-76
[PubMed:15378759] [WorldCat.org] [DOI] (P p)

D B Mirel, W F Estacio, M Mathieu, E Olmsted, J Ramirez, L M Márquez-Magaña
Environmental regulation of Bacillus subtilis sigma(D)-dependent gene expression.
J Bacteriol: 2000, 182(11);3055-62
[PubMed:10809682] [WorldCat.org] [DOI] (P p)

T Caramori, A Galizzi
The UP element of the promoter for the flagellin gene, hag, stimulates transcription from both SigD- and SigA-dependent promoters in Bacillus subtilis.
Mol Gen Genet: 1998, 258(4);385-8
[PubMed:9648743] [WorldCat.org] [DOI] (P p)

Y F Chen, J D Helmann
DNA-melting at the Bacillus subtilis flagellin promoter nucleates near -10 and expands unidirectionally.
J Mol Biol: 1997, 267(1);47-59
[PubMed:9096206] [WorldCat.org] [DOI] (P p)

K Fredrick, T Caramori, Y F Chen, A Galizzi, J D Helmann
Promoter architecture in the flagellar regulon of Bacillus subtilis: high-level expression of flagellin by the sigma D RNA polymerase requires an upstream promoter element.
Proc Natl Acad Sci U S A: 1995, 92(7);2582-6
[PubMed:7708689] [WorldCat.org] [DOI] (P p)

D B Mirel, M J Chamberlin
The Bacillus subtilis flagellin gene (hag) is transcribed by the sigma 28 form of RNA polymerase.
J Bacteriol: 1989, 171(6);3095-101
[PubMed:2498284] [WorldCat.org] [DOI] (P p)