AbiV, a novel antiphage abortive infection mechanism on the chromosome of Lactococcus lactis subsp. cremoris MG1363.
Insertional mutagenesis with pGhost9::ISS1 resulted in independent insertions in a 350-bp region of the chromosome of Lactococcus lactis subsp. cremoris MG1363 that conferred phage resistance to the integrants. The orientation and location of the insertions suggested that the phage resistance phenotype was caused by a chromosomal gene turned on by a promoter from the inserted construct. Reverse transcription-PCR analysis confirmed that there were higher levels of transcription of a downstream open reading frame (ORF) in the phage-resistant integrants than in the phage-sensitive strain L. lactis MG1363. This gene was also found to confer phage resistance to L. lactis MG1363 when it was cloned into an expression vector. A subsequent frameshift mutation in the ORF completely eliminated the phage resistance phenotype, confirming that the ORF was necessary for phage resistance. This ORF provided resistance against virulent lactococcal phages belonging to the 936 and c2 species with an efficiency of plaquing of 10(-4), but it did not protect against members of the P335 species. A high level of expression of the ORF did not affect the cellular growth rate. Assays for phage adsorption, DNA ejection, restriction/modification activity, plaque size, phage DNA replication, and cell survival showed that the ORF encoded an abortive infection (Abi) mechanism. Sequence analysis revealed a deduced protein consisting of 201 amino acids which, in its native state, probably forms a dimer in the cytosol. Similarity searches revealed no homology to other phage resistance mechanisms, and thus, this novel Abi mechanism was designated AbiV. The mode of action of AbiV is unknown, but the activity of AbiV prevented cleavage of the replicated phage DNA of 936-like phages.
Haaber
Jakob
J
Center for Systems Microbiology, DTU Biosys, Technical University of Denmark, Lyngby, Denmark.
Moineau
Sylvain
S
Fortier
Louis-Charles
LC
Hammer
Karin
K
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D001435:Bacteriophages / Q000254:growth & development* / Q000276:immunology*; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D019281:Dimerization; D016368:Frameshift Mutation; D018628:Gene Dosage; D020869:Gene Expression Profiling; D023061:Gene Order; D013294:Lactococcus lactis / Q000502:physiology* / Q000821:virology*; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D012329:RNA, Bacterial / Q000096:biosynthesis; D012333:RNA, Messenger / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D017906:Siphoviridae / Q000254:growth & development / Q000276:immunology; D010948:Viral Plaque Assay
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D012329:RNA, Bacterial; D012333:RNA, Messenger
Antonie Van Leeuwenhoek. 1999 Jul-Nov;76(1-4):89-113
10532374
J Bacteriol. 1995 Jul;177(13):3818-23
7601848
Appl Environ Microbiol. 1993 Jan;59(1):208-12
16348844
Mol Microbiol. 2000 Feb;35(3):517-28
10672175
Appl Environ Microbiol. 1991 Jan;57(1):283-8
16348396
Virology. 1978 Mar;85(1):118-28
347693
Microbiology (Reading). 2002 Apr;148(Pt 4):985-1001
11932445
Gene. 1998 May 28;212(1):5-11
9661658
Syst Appl Microbiol. 1983;4(3):413-23
23194740
Genome Res. 2001 May;11(5):731-53
11337471
Antonie Van Leeuwenhoek. 1999 Jul-Nov;76(1-4):377-82
10532393
Appl Environ Microbiol. 1996 Sep;62(9):3075-82
8795193
Appl Environ Microbiol. 1994 Jun;60(6):1832-41
16349277
J Biotechnol. 1999 Jan 22;67(2-3):135-49
9990732
Virology. 2002 May 10;296(2):308-20
12069529
Virology. 2000 Sep 30;275(2):267-77
10998327
J Bacteriol. 1992 Sep;174(17):5633-8
1324906
Appl Environ Microbiol. 1980 Sep;40(3):500-6
16345629
J Bacteriol. 1983 Apr;154(1):1-9
6403500
Virology. 2008 Apr 10;373(2):298-309
18191977
Appl Environ Microbiol. 1987 Oct;53(10):2452-7
2447829
Appl Environ Microbiol. 1991 May;57(5):1346-53
16348479
J Bacteriol. 1995 Jul;177(13):3824-9
7601849
FEMS Microbiol Lett. 2006 Aug;261(2):253-61
16907729
J Bacteriol. 1994 Nov;176(21):6457-63
7961396
J Dairy Sci. 1998 Jun;81(6):1483-5
9684156
Appl Environ Microbiol. 1995 Jun;61(6):2193-202
7793939
Appl Environ Microbiol. 2004 Oct;70(10):5825-32
15466520
Mol Microbiol. 1997 Oct;26(1):49-64
9383189
J Bacteriol. 1996 Feb;178(3):931-5
8550537
Appl Environ Microbiol. 1989 Dec;55(12):3119-23
16348073
J Bacteriol. 1998 Jun;180(12):3174-80
9620968
Proc Natl Acad Sci U S A. 1987 Jun;84(12):4171-5
2438693
Appl Environ Microbiol. 1995 Dec;61(12):4321-8
8534099
Appl Environ Microbiol. 1999 Nov;65(11):4793-8
10543788
FEMS Microbiol Lett. 1996 Sep 1;142(2-3):295-9
8810513
J Bacteriol. 2007 Feb;189(4):1482-7
17041060
Appl Environ Microbiol. 1990 Jan;56(1):202-9
16348092
Appl Environ Microbiol. 1997 Apr;63(4):1274-83
9097424
Appl Environ Microbiol. 1996 Feb;62(2):676-9
16535245
Microbiology (Reading). 2000 Feb;146 ( Pt 2):445-453
10708383
Appl Microbiol. 1975 Jun;29(6):807-13
16350018
Appl Environ Microbiol. 1991 Dec;57(12):3547-51
1664711
Appl Environ Microbiol. 1993 Dec;59(12):4363-6
16349136
Appl Environ Microbiol. 2000 Sep;66(9):3790-7
10966392
Appl Environ Microbiol. 2001 Nov;67(11):5225-32
11679349
J Bacteriol. 2004 Feb;186(3):713-21
14729697
J Dairy Sci. 2000 Jul;83(7):1448-50
10908051
FEMS Microbiol Lett. 1998 Feb 15;159(2):331-6
9503629
Nucleic Acids Res. 1997 Sep 1;25(17):3389-402
9254694
Appl Environ Microbiol. 1999 Jan;65(1):330-5
9872803
FEMS Microbiol Lett. 2002 Sep 10;214(2):283-7
12351244
J Bacteriol. 1997 May;179(9):2949-57
9139913
J Bacteriol. 2002 Nov;184(22):6325-32
12399502
J Bacteriol. 2007 Feb;189(4):1417-25
17012400
Appl Environ Microbiol. 1994 Apr;60(4):1129-36
16349225
Curr Opin Microbiol. 2005 Aug;8(4):473-9
15979388
Microbiology (Reading). 1994 Nov;140 ( Pt 11):3061-9
7812447
Appl Environ Microbiol. 1990 Jul;56(7):2255-8
2389939
J Bacteriol. 2007 Apr;189(8):3256-70
17307855
Appl Environ Microbiol. 1995 May;61(5):2023-6
7646042
Mol Gen Genet. 1996 Nov 27;253(1-2):217-24
9003306
Appl Environ Microbiol. 1998 Dec;64(12):4748-56
9835558
J Bacteriol. 1986 Sep;167(3):905-19
3017919
Antonie Van Leeuwenhoek. 2002 Aug;82(1-4):303-21
12369198
J Bacteriol. 2001 May;183(9):2785-94
11292797