NtcA from Microcystis aeruginosa PCC 7806 is autoregulatory and binds to the microcystin promoter.
NtcA is a transcription factor that has been found in a diverse range of cyanobacteria. This nitrogen-controlled factor was focused on as a key component in the yet-to-be-deciphered regulatory network controlling microcystin production. Adaptor-mediated PCR was utilized to isolate the ntcA gene from Microcystis aeruginosa PCC 7806. This gene was cloned, and the recombinant (His-tagged) protein was overexpressed and purified for use in mobility shift assays to analyze NtcA binding to putative sites identified in the microcystin mcyA/D promoter region. Autoregulation of NtcA in M. aeruginosa was shown via NtcA binding in the upstream ntcA promoter region. The observation of binding of NtcA to the mcyA/D promoter region has direct relevance for the regulation of microcystin biosynthesis, as transcription of the mcyABCDEFGHIJ gene cluster appears to be under direct control of nitrogen.
Ginn
H P
HP
Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales 2052, Australia.
Pearson
L A
LA
Neilan
B A
BA
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial; D052998:Microcystins / Q000096:biosynthesis*; D046931:Microcystis / Q000235:genetics / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D052998:Microcystins; D011994:Recombinant Proteins; D014157:Transcription Factors; C078588:microcystin; D009584:Nitrogen
Plant Physiol. 1996 Jul;111(3):839-48
8754684
J Bacteriol. 2001 Jan;183(2):411-25
11133933
Appl Environ Microbiol. 1995 Feb;61(2):797-800
7574617
J Appl Bacteriol. 1992 Jun;72(6):445-59
1644701
Toxicol Appl Pharmacol. 1995 Oct;134(2):264-72
7570603
FEMS Microbiol Lett. 2001 Mar 15;196(2):207-14
11267781
Appl Environ Microbiol. 1995 Nov;61(11):3875-83
8526499
J Bacteriol. 1993 Jul;175(13):4025-35
8391534
Methods. 2001 Dec;25(4):402-8
11846609
Plant Physiol. 1998 May;117(1):129-39
9576782
FEMS Microbiol Lett. 2004 Jul 1;236(1):47-52
15212789
Appl Environ Microbiol. 1999 Feb;65(2):737-9
9925609
Mol Microbiol. 2002 Jun;44(5):1377-85
12068814
Biochim Biophys Acta. 2004 Aug 12;1679(2):156-63
15297148
EMBO J. 1994 Jun 15;13(12):2862-9
8026471
J Bacteriol. 1990 Feb;172(2):643-7
1967601
J Biol Chem. 1995 Sep 29;270(39):22882-9
7559423
J Bacteriol. 1998 Apr;180(7):1878-86
9537388
Mol Gen Genet. 1993 Jan;236(2-3):193-202
8437564
Microbiology (Reading). 2001 Nov;147(Pt 11):3113-9
11700361
Mol Microbiol. 1992 Jul;6(13):1853-9
1630321
Biochem J. 1997 Oct 15;327 ( Pt 2):513-7
9359424
J Mol Biol. 2000 Aug 25;301(4):783-93
10966785
Protein Expr Purif. 1999 Dec;17(3):351-7
10600452
Arch Microbiol. 1995 Jan;163(1):7-15
7710323
J Bacteriol. 1996 Feb;178(3):922-6
8550535
J Bacteriol. 1993 Sep;175(17):5710-3
8366058
Lancet. 1998 Jul 4;352(9121):21-6
9800741
Chem Biol. 2000 Oct;7(10):753-64
11033079
Appl Environ Microbiol. 2000 Aug;66(8):3387-92
10919796
Proc Natl Acad Sci U S A. 1998 Sep 1;95(18):11008-13
9724820
Appl Environ Microbiol. 2004 Nov;70(11):6353-62
15528492