Abstract
A protein of cyAbrB family of regulators, Alr0946 (CalA), was identified by DNA affinity chromatography using DNA fragments from the promoter region of ftsZ in Anabaena/Nostoc sp. PCC 7120. Electrophoretic mobility shift assays showed that the protein could bind to upstream regions of ftsZ (as biotin-labeled DNA fragments) and that the binding was inhibited by the same DNA fragments and a fragment upstream of hetC but not by 3 fragments upstream of glnA, hetR or ntcA. Both transcriptional fusion with gfp (green fluorescence protein) and Western blot analysis indicated that the expression of calA was down-regulated in heterocysts relative to vegetative cells. An insertional mutant of calA could not be completely segregated. Over-expression of calA in Anabaena/Nostoc caused no change in growth, heterocyst differentiation or expression of ftsZ. CalA as a DNA-binding protein is essential for the growth of Anabaena/Nostoc and may be required for the expression of ftsZ in vegetative cells.
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References
Adams DW, Errington J (2009) Bacterial cell division: assembly, maintenance and disassembly of the Z ring. Nat Rev Microbiol 7:642–653
Agervald Å, Zhang X, Stensjö K, Devine E, Lindblad P (2010) CalA, a cyanobacterial AbrB protein, interacts with the upstream region of hypC and acts as a repressor of its transcription in the cyanobacterium Nostoc sp. strain PCC 7120. Appl Environ Microbiol 76:880–890
Black TA, Wolk CP (1994) Analysis of a Het- mutation in Anabaena sp. strain PCC 7120 implicates a secondary metabolite in the regulation of heterocyst spacing. J Bacteriol 176:2282–2292
Black TA, Cai Y, Wolk CP (1993) Spatial expression and autoregulation of hetR, a gene involved in the control of heterocyst development in Anabaena. Mol Microbiol 9:77–84
Buikema WJ, Haselkorn R (2001) Expression of the Anabaena hetR gene from a copper-regulated promoter leads to heterocyst differentiation under repressing conditions. Proc Natl Acad Sci USA 98:2729–2734
Cai Y, Wolk CP (1990) Use of a conditionally lethal gene in Anabaena sp. strain PCC 7120 to select for double recombinants and to entrap insertion sequences. J Bacteriol 172:3138–3145
Chu F, Kearns DB, McLoon A, Chai YR, Kolter R, Losick R (2008) A novel regulatory protein governing biofilm formation in Bacillus subtilis. Mol Microbiol 68:1117–1127
Elhai J, Wolk CP (1988a) Conjugal transfer of DNA to cyanobacteria. Methods Enzymol 167:747–754
Elhai J, Wolk CP (1988b) A versatile class of positive-selection vectors based on the nonviability of palindrome-containing plasmids that allows cloning into long polylinkers. Gene 68:119–138
Fisher R, Tuli R, Haselkorn R (1981) A cloned cyanobacterial gene for glutamine synthetase functions in Escherichia coli, but the enzyme is not adenylylated. Proc Natl Acad Sci USA 78:3393–3397
Ghassemian M, Wong B, Ferreira F, Markley JL, Straus NA (1994) Cloning, sequencing and transcriptional studies of the genes for cytochrome c-553 and plastocyanin from Anabaena sp. PCC 7120. Microbiology 140:1151–1159
Herrero A, Muro-Pastor AM, Valladares A, Flores E (2004) Cellular differentiation and the NtcA transcription factor in filamentous cyanobacteria. FEMS Microbiol Rev 28:469–487
Huang X, Dong Y, Zhao J (2004) HetR homodimer is a DNA-binding protein required for heterocyst differentiation, and the DNA-binding activity is inhibited by PatS. Proc Natl Acad Sci USA 101:4848–4853
Ishii A, Hihara Y (2008) An AbrB-like transcriptional regulator, Sll0822, is essential for the activation of nitrogen-regulated genes in Synechocystis sp. PCC 6803. Plant Physiol 148:660–670
Khudyakov I, Wolk CP (1997) hetC, a gene coding for a protein similar to bacterial ABC protein exporters, is involved in early regulation of heterocyst differentiation in Anabaena sp. strain PCC 7120. J Bacteriol 179:6971–6978
Klint J, Rasmussen U, Bergman B (2007) FtsZ may have dual roles in the filamentous cyanobacterium Nostoc/Anabaena sp. strain PCC 7120. J Plant Physiol 164:11–18
Kuhn I, Peng L, Bedu S, Zhang CC (2000) Developmental regulation of the cell division protein FtsZ in Anabaena sp. strain PCC 7120, a cyanobacterium capable of terminal differentiation. J Bacteriol 182:4640–4643
Laurent S, Chen H, Bédu S, Ziarelli F, Peng L, Zhang CC (2005) Nonmetabolizable analogue of 2-oxoglutarate elicits heterocyst differentiation under repressive conditions in Anabaena sp. PCC 7120. Proc Natl Acad Sci USA 102:907–1012
Lieman-Hurwitz J, Haimovich M, Shalev-Malul G, Ishii A, Hihara Y, Gaathon A, Lebendiker M, Kaplan A (2009) A cyanobacterial AbrB-like protein affects the apparent photosynthetic affinity for CO2 by modulating low-CO2-induced gene expression. Environ Microbiol 11:927–936
Muro-Pastor AM, Valladares A, Flores E, Herrero A (1999) The hetC gene is a direct target of the NtcA transcriptional regulator in cyanobacterial heterocyst development. J Bacteriol 181:6664–6669
Oliveira P, Lindblad P (2008) An AbrB-Like protein regulates the expression of the bidirectional hydrogenase in Synechocystis sp. strain PCC 6803. J Bacteriol 190:1011–1019
Phillips ZEV, Strauch MA (2002) Bacillus subtilis sporulation and stationary phase gene expression. Cell Mol Life Sci 59:392–402
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61
Sakr S, Jeanjean R, Zhang CC, Arcondeguy T (2006) Inhibition of cell division suppresses heterocyst development in Anabaena sp. strain PCC 7120. J Bacteriol 2:1396–1404
Shalev-Malul G, Lieman-Hurwitz J, Viner-Mozzini Y, Sukenik A, Gaathon A, Lebendiker M, Kaplan A (2008) An AbrB-like protein might be involved in the regulation of cylindrospermopsin production by Aphanizomenon ovalisporum. Environ Microbiol 10:988–999
Shi Y, Zhao W, Zhang W, Ye Z, Zhao J (2006) Regulation of intracellular free calcium concentration during heterocyst differentiation by HetR and NtcA in Anabaena sp. PCC 7120. Proc Natl Acad Sci USA 103:11334–11339
Strauch MA, Hoch JA (1993) Transition-state regulators: sentinels of Bacillus subtilis post-exponential gene expression. Mol Microbiol 7:337–342
Valladares A, Muro-Pastor AM, Herrero A, Flores E (2004) The NtcA-dependent P1 promoter is utilized for glnA expression in N2-fixing heterocysts of Anabaena sp. strain PCC 7120. J Bacteriol 186:7337–7343
Wang Y, Xu X (2005) Regulation by hetC of genes required for heterocyst differentiation and cell division in Anabaena sp. strain PCC 7120. J Bacteriol 187:8489–8493
Wolk CP, Ernst A, Elhai J (1994) Heterocyst metabolism and development. In: Bryant DA (ed) The molecular biology of cyanobacteria. Kluwer Academic Publishers, Dordrecht, pp 769–823
Xu K, Strauch MA (1996) In vitro selection of optimal AbrB-binding sites: comparison to known in vivo sites indicates flexibility in AbrB binding and recognition of three-dimensional DNA structures. Mol Microbiol 19:145–158
Xu X, Wolk CP (2001) Role for hetC in the transition to a nondividing state during heterocyst differentiation in Anabaena sp. J Bacteriol 183:393–396
Xu X, Elhai J, Wolk CP (2008) Transcriptional and developmental responses by Anabaena to deprivation of fixed nitrogen. In: Herrero T, Flores E (eds) Cyanobacteria: molecular biology, genomics and evolution. Horizon Scientific Press, Norwich, pp 383–422
Zhang W, Du Y, Khudyakov I, Fan Q, Gao H, Ning D, Wolk CP, Xu X (2007) A gene cluster that regulates both heterocyst differentiation and pattern formation in Anabaena sp. strain PCC 7120. Mol Microbiol 66:1429–1443
Zhou R, Wei X, Jiang N, Li H, Dong Y, His KL, Zhao J (1998) Evidence that HetR protein is an unusual serine-type protease. Proc Natl Acad Sci USA 95:4959–4963
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This work was supported by the National Natural Science Foundation of China under grants 30570022 and 30770049.
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Communicated by Jack Meeks.
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He, D., Xu, X. CalA, a cyAbrB protein, binds to the upstream region of ftsZ and is down-regulated in heterocysts in Anabaena sp. PCC 7120. Arch Microbiol 192, 461–469 (2010). https://doi.org/10.1007/s00203-010-0573-9
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DOI: https://doi.org/10.1007/s00203-010-0573-9