YneA, an SOS-induced inhibitor of cell division in Bacillus subtilis, is regulated posttranslationally and requires the transmembrane region for activity.
Cell viability depends on the stable transmission of genetic information to each successive generation. Therefore, in the event of intrinsic or extrinsic DNA damage, it is important that cell division be delayed until DNA repair has been completed. In Bacillus subtilis, this is accomplished in part by YneA, an inhibitor of division that is induced as part of the SOS response. We sought to gain insight into the mechanism by which YneA blocks cell division and the processes involved in shutting off YneA activity. Our data suggest that YneA is able to inhibit daughter cell separation as well as septum formation. YneA contains a LysM peptidoglycan binding domain and is predicted to be exported. We established that the YneA signal peptide is rapidly cleaved, resulting in secretion of YneA into the medium. Mutations within YneA affect both the rate of signal sequence cleavage and the activity of YneA. YneA does not stably associate with the cell wall and is rapidly degraded by extracellular proteases. Based on these results, we hypothesize that exported YneA is active prior to signal peptide cleavage and that proteolysis contributes to the inactivation of YneA. Finally, we identified mutations in the transmembrane segment of YneA that abolish the ability of YneA to inhibit cell division, while having little or no effect on YneA export or stability. These data suggest that protein-protein interactions mediated by the transmembrane region may be required for YneA activity.
Mo
Allison H
AH
Department of Biology, Stanford University, 371 Serra Mall, Stanford, CA 94305-5020, USA.
Burkholder
William F
WF
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002455:Cell Division; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005838:Genotype; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D020664:Son of Sevenless Proteins / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D003598:Cytoskeletal Proteins; C071525:FtsZ protein, Bacteria; D020664:Son of Sevenless Proteins
Mol Microbiol. 1998 Mar;27(5):899-914
9535081
J Mol Biol. 2004 Jul 16;340(4):783-95
15223320
Microbiol Mol Biol Rev. 2000 Sep;64(3):515-47
10974125
Mol Microbiol. 2006 Oct;62(1):132-47
16942606
Annu Rev Cell Dev Biol. 2003;19:565-87
14570582
J Bacteriol. 1996 Sep;178(17):5080-5
8752322
Microbiology (Reading). 2000 Feb;146 ( Pt 2):249-262
10708363
J Bacteriol. 1983 Jan;153(1):169-75
6336731
J Bacteriol. 2006 Nov;188(21):7396-404
16936019
Int J Biol Sci. 2008 Sep 23;4(6):338-44
18825275
Proc Natl Acad Sci U S A. 1986 Jul;83(14):5204-8
3088571
J Mol Biol. 2000 Jun 16;299(4):1113-9
10843862
J Bacteriol. 2003 Aug;185(16):4861-71
12897006
Mol Microbiol. 2008 May;68(4):838-47
18430080
Biochimie. 1997 Oct;79(9-10):549-54
9466691
Microbiol Mol Biol Rev. 2004 Jun;68(2):207-33
15187182
Mol Microbiol. 1999 Jul;33(1):33-40
10411721
J Bacteriol. 2003 Nov;185(22):6666-77
14594841
Microbiology (Reading). 2000 Jan;146 ( Pt 1):65-75
10658653
Annu Rev Genet. 2000;34:479-497
11092836
Appl Environ Microbiol. 1999 Jul;65(7):2934-41
10388686
J Bacteriol. 1998 Feb;180(3):749-52
9457885
Proteomics. 2002 May;2(5):591-602
11987133
Acta Biochim Pol. 2001;48(3):599-610
11833768
Microbiology (Reading). 2006 Oct;152(Pt 10):2867-2874
17005968
Proc Natl Acad Sci U S A. 1984 Jul;81(14):4490-4
6087326
Mol Microbiol. 2000 Apr;36(2):278-89
10792716
Proc Natl Acad Sci U S A. 2003 Jun 24;100(13):7889-94
12808143
J Gen Microbiol. 1977 Nov;103(1):201-5
412916
Mol Microbiol. 2000 Jan;35(2):299-311
10652091
Mol Microbiol. 2008 Feb;67(3):597-608
18086211
J Bacteriol. 1998 Aug;180(15):3946-53
9683493
J Bacteriol. 1991 Apr;173(7):2278-82
1901054
J Bacteriol. 2006 Aug;188(15):5541-50
16855244
J Biol Chem. 1997 Oct 10;272(41):25983-92
9325333
FEBS Lett. 1994 Jun 6;346(1):17-20
8206151
J Bacteriol. 1995 Dec;177(24):7231-7
8522532
Biochemistry. 2003 Jan 21;42(2):257-64
12525152
Bioessays. 1995 Jan;17(1):63-70
7702595
J Bacteriol. 1991 Jan;173(1):276-82
1898923
Mol Microbiol. 2006 Jun;60(6):1490-508
16796683
Basic Life Sci. 1975;5A:355-67
1103845
Mol Microbiol. 2008 Mar;67(5):1143-55
18208530
J Bacteriol. 2006 Aug;188(16):6034-8
16885473
Mol Microbiol. 2006 Dec;62(5):1228-38
17042786
Curr Biol. 2005 Jul 12;15(13):R514-26
16005287
J Bacteriol. 1973 Sep;115(3):1063-70
4199504
J Bacteriol. 1996 Jan;178(1):156-62
8550410
Microbiology (Reading). 1998 Jan;144 ( Pt 1):73-82
9537764
J Bacteriol. 2005 Nov;187(22):7655-66
16267290
Appl Environ Microbiol. 2010 Mar;76(6):1992-5
20097825
Gene. 2001 Feb 21;264(2):289-97
11250085
Mol Genet Genomics. 2003 Mar;268(6):761-70
12655402
Microbiol Mol Biol Rev. 2003 Mar;67(1):52-65, table of contents
12626683
Mol Microbiol. 2003 Feb;47(4):1113-22
12581363
Proc Natl Acad Sci U S A. 1983 Jan;80(2):358-62
6300834
Microbiology (Reading). 2010 Feb;156(Pt 2):374-384
19892760
Cell. 1982 May;29(1):11-22
7049397