Isolation of a prokaryotic metallothionein locus and analysis of transcriptional control by trace metal ions

Molecular Microbiology - Tập 7 Số 2 - Trang 177-187 - 1993
James W. Huckle1, Andrew P. Morby1, Jim Cavet1, Nigel J. Robinson1
1Department of Biological Sciences, University of Durham, South Road, Durham, DH1 3LE, UK

Tóm tắt

SummaryIn eukaryotes, metallothioneins (MTs) are involved in cellular responses to elevated concentrations of certain metal ions. We report the isolation and analysis of a prokaryotic MT locus from Synechococcus PCC 7942. The MT locus (smt) includes smtA, which encodes a class II MT, and a divergently transcribed gene, smtB. The sites of transcription initiation of both genes have been mapped and features within the smt operator‐promoter region identified. Elevated concentrations of the ionic species of Cd, Co, Cr, Cu, Hg, Ni, Pb and Zn elicited an increase in the abundance of smtA transcripts. There was no detectable effect of elevated metal (Cd) on smtA transcript stability. Sequences upstream of smtA, fused to a promoterless lacZ gene, conferred metal‐dependent β‐galactosidase activity in Synechococcus PCC 7942 (strain R2‐PIM8). At maximum permissive concentrations, Zn was the most potent elicitor in vivo, followed by Cu and Cd with slight induction by Co and Ni. The deduced SmtB polypeptide has similarity to the ArsR and CadC proteins involved in resistance to arsenate/arsenite/antimonite and to Cd, contains a predicted helix‐turn‐helix DNA‐binding motif and is shown to be a repressor of transcription from the smtA operator‐promoter.

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Tài liệu tham khảo

10.1016/0378-1119(84)90197-5

10.1111/j.1529-8817.1968.tb04667.x

10.1038/355087a0

10.1128/MMBR.51.4.509-518.1987

Beck CF., 1988, Divergent promoters, a common form of gene organization, Microbiol Rev, 5, 318, 10.1128/mr.52.3.318-326.1988

10.1016/0147-619X(92)90005-U

10.1073/pnas.81.11.3332

10.1016/0162-0134(91)84121-O

Dodd I.B., 1990, Improved detection of helix‐turn‐helix DNA‐binding motifs in protein sequences, Nucl Acids Res, 18, 5019, 10.1093/nar/18.17.5019

Dzelzkalns V.A., 1988, Plant Molecular Biology: A Practical Approach, 277

10.1016/0003-2697(83)90418-9

10.1111/j.1365-2958.1993.tb01110.x

Hamer D.H., 1986, Metallothioneins, Ann Pev Biochem, 55, 913, 10.1146/annurev.bi.55.070186.004405

10.1111/j.1365-2958.1990.tb00538.x

10.1128/jb.174.11.3684-3694.1992

10.1016/0076-6879(91)05145-L

10.1073/pnas.81.2.337

10.1016/0076-6879(91)05078-A

10.1002/j.1537-2197.1955.tb11120.x

10.1016/0147-619X(83)90068-9

10.1093/nar/19.15.4225

10.1093/nar/17.14.5517

Mack Ivey D., 1992, The cadC gene products of alkaphilic Bacillus firmus OF4 partially restores Na+ resistance to an Escherichia coli strain lacking an Na+/H+ antiporter (NhaA), J Bacteriol, 174, 4878, 10.1128/jb.174.15.4878-4884.1992

10.1016/0378-1119(88)90608-7

Miller J.H., 1972, Experiments in Molecular Genetics, 351

10.1042/bj2510691

10.1007/978-3-0348-6784-9_4

Riordan J.F., 1991, Metallobiochemistry part B, metallothionein and related molecules, Meth Enzymol, 205

10.1016/0168-9452(88)90098-2

10.1098/rspb.1990.0130

10.1128/jb.174.11.3676-3683.1992

Rouch D.A., 1989, Metal Ion Homeostasis: Molecular Biology and Chemistry, 469

Sambrook J., 1989, Molecular cloning: A Laboratory Manual

10.1093/nar/18.3.619

Sanger F., 1977, DNA sequencing with chain‐terminating inhibitors, Proc Natl Acad Sci USA, 7, 5463, 10.1073/pnas.74.12.5463

10.1016/0378-1119(90)90437-V

10.1080/00071618200650021

10.1016/0014-5793(92)80509-F

10.1146/annurev.mi.42.100188.003441

10.1128/jb.146.3.983-996.1981

10.1016/0076-6879(91)05077-9

Wealand J.L., 1989, Changes in gene expression during nitrogen starvation in Anabaena variabilis ATCC 29413, J Bacteriol, 171, 1309, 10.1128/jb.171.3.1309-1313.1989

10.1111/j.1365-2958.1991.tb00779.x

10.1128/jb.173.23.7636-7642.1991

10.1016/0014-5793(91)80175-3