Envelope stress is a trigger of CRISPR RNA-mediated DNA silencing in Escherichia coli.
A widespread feature in the genomes of most bacteria and archaea is an array of clustered, regularly interspaced short palindromic repeats (CRISPRs) that, together with a group of CRISPR-associated (Cas) proteins, mediate immunity against invasive nucleic acids such as plasmids and viruses. Here, the CRISPR-Cas system was activated in cells expressing a plasmid-encoded protein that was targeted to the twin-arginine translocation (Tat) pathway. Expression of this Tat substrate resulted in upregulation of the Cas enzymes and subsequent silencing of the encoding plasmid in a manner that required the BaeSR two-component regulatory system, which is known to respond to extracytoplasmic stress. Furthermore, we confirm that the CasCDE enzymes form a stable ternary complex and appear to function as the catalytic core of the Cas system to process CRISPR RNA into its mature form. Taken together, our results indicate that the CRISPR-Cas system targets DNA directly as part of a defence mechanism in bacteria that is overlapping with but not limited to phage infection.
School of Chemical and Biomolecular Engineering Department of Microbiology Macromolecular Diffraction Facility at CHESS Department of Molecular Biology and Genetics Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853 USA.
Ritsdeliz
R
Perez-Rodriguez
Charles
C
Haitjema
Qingqiu
Q
Huang
Ki Hyun
KH
Nam
Sarah
S
Bernardis
Ailong
A
Ke
Matthew P
MP
DeLisa
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000378:metabolism; D020868:Gene Silencing; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins / Q000378:metabolism; D055029:Inverted Repeat Sequences / Q000235:genetics*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011387:Prokaryotic Cells / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012329:RNA, Bacterial / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics*
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase; D012260:Ribonucleases
Mol Microbiol. 2001 Jul;41(1):241-6
11454216
Cell. 2009 Feb 20;136(4):615-28
19239884
Science. 2008 Dec 19;322(5909):1843-5
19095942
J Biotechnol. 2008 Jan 20;133(2):196-200
17884222
RNA. 2008 Dec;14(12):2572-9
18971321
PLoS Comput Biol. 2005 Nov;1(6):e60
16292354
Mol Microbiol. 2005 Feb;55(4):1113-26
15686558
Mol Microbiol. 2009 Apr;72(1):259-72
19239620
J Mol Biol. 2008 Jun 27;380(1):51-66
18511069
Nat Biotechnol. 2004 Dec;22(12):1554-8
15543133
J Bacteriol. 2008 Feb;190(4):1390-400
18065545
Proc Natl Acad Sci U S A. 2002 May 28;99(11):7536-41
12032318
J Bacteriol. 2002 Aug;184(15):4168-76
12107134
Microbiology (Reading). 2010 May;156(5):1351-1361
28206910
Lab Chip. 2006 Mar;6(3):381-8
16511621
Science. 2010 Sep 10;329(5997):1355-8
20829488
Mol Microbiol. 2002 Sep;45(6):1599-611
12354228
Mol Syst Biol. 2006;2:2006.0008
16738554
Appl Environ Microbiol. 2005 Dec;71(12):8451-9
16332834
Mol Microbiol. 1997 Apr;24(2):255-61
9159513
Nucleic Acids Res. 2007 Jul;35(Web Server issue):W52-7
17537822
Science. 2007 Mar 23;315(5819):1709-12
17379808
J Bacteriol. 1989 May;171(5):2581-90
2540151
Adv Exp Med Biol. 2008;631:80-110
18792683
Nat Rev Microbiol. 2008 Mar;6(3):181-6
18157154
Mol Microbiol. 2010 Sep;77(6):1380-93
20659289
Anal Biochem. 1997 Dec 1;254(1):150-2
9398359
Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5
10829079
J Biol Chem. 2002 Aug 16;277(33):29825-31
12021272
Mol Microbiol. 2010 Mar;75(6):1495-512
20132443
J Mol Biol. 2006 Feb 03;355(5):957-67
16343541
Nucleic Acids Res. 2008 Jul 1;36(Web Server issue):W145-8
18442988
Proc Natl Acad Sci U S A. 2003 May 13;100(10):6115-20
12721369
Biol Direct. 2006 Mar 16;1:7
16545108
Genome Biol. 2007;8(4):R61
17442114
Science. 2008 Aug 15;321(5891):960-4
18703739
Archaea. 2006 Aug;2(1):59-72
16877322
Mol Microbiol. 2001 Apr;40(2):323-31
11309116
J Biol Chem. 2005 Dec 30;280(52):42723-30
16263723
Genes Dev. 2008 Dec 15;22(24):3489-96
19141480
Nature. 2010 Jan 28;463(7280):568-71
20072129
Science. 2004 Sep 3;305(5689):1437-41
15284456
J Mol Biol. 2007 Mar 30;367(3):715-30
17280684
Cell. 2009 Nov 25;139(5):945-56
19945378
J Bacteriol. 2004 Jan;186(2):366-73
14702305