Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The mercury resistance (mer) operon of the gram-negative transposon Tn21 encodes not only a mercuric reductase and regulatory genes but also two inner membrane proteins (MerT and MerC) and a periplasmic protein (MerP). Although the merT, merP, and merC genes have been implicated in Hg(II) transport, the individual roles of these genes have not been established. We created in vitro precise deletion and frameshift mutations that eliminated each of the genes singly and in combination. Our results show that both merT and merP are required for Hg(II) binding but that merC is not. Both merT and merP are required for full expression of Hg(II) resistance, but loss of merP is less deleterious than loss of merT. Furthermore, mutations eliminating both merT and merP decrease resistance more than the single mutations do. In contrast, mutating merC had no effect on Hg(II) resistance. Both the merT and merP mutations increase the threshold Hg(II) concentration for induction of merA-lacZ transcriptional fusions and cause an increase in the maximal expression level. In contrast, the merC mutation had little effect on the threshold inducing concentration of Hg(II) but decreased the level of expression. Our results show that merT and merP alone are sufficient to specify a mercury transport system. The role of merC remains obscure.

Bibliography

Hamlett, N. V., Landale, E. C., Davis, B. H., & Summers, A. O. (1992). Roles of the Tn21 merT, merP, and merC gene products in mercury resistance and mercury binding. Journal of Bacteriology, 174(20), 6377–6385.

Authors 4
  1. N V Hamlett (first)
  2. E C Landale (additional)
  3. B H Davis (additional)
  4. A O Summers (additional)
References 51 Referenced 122
  1. Barrineau , P. , and A. 0. Summers . 1983 . A second positive regulatory function in the mer (mercury resistance) operon . Gene 25 : 209 - 211 . (10.1016/0378-1119(83)90225-1) / Gene / A second positive regulatory function in the mer (mercury resistance) operon by Barrineau P. (1983)
  2. Barrineau , P. , P. Gilbert , W. J. Jackson , C. S. Jones , A. 0. Summers , and S. Wisdom . 1984 . The DNA sequence of the mercury resistance operon of the IncFII plasmid NR1 . J. Mol. Appl. Genet. 2 : 601 - 619 . / J. Mol. Appl. Genet. / The DNA sequence of the mercury resistance operon of the IncFII plasmid NR1 by Barrineau P. (1984)
  3. Brown , N. L. , J. Camakaris , B. T. 0. Lee , T. Williams , A. P. Morby , J. Parkhiil , and D. A. Rouch . 1991 . Bacterial resistances to mercury and copper . J. Cell Biochem. 46 : 106 - 114 . (10.1002/jcb.240460204) / J. Cell Biochem. / Bacterial resistances to mercury and copper by Brown N. L. (1991)
  4. Brown , N. L. , S. J. Ford , R. D. Pridmore , and D. C. Fritzinger . 1983 . Nucleotide sequence of a gene from the Pseudomonas transposon Tn 501 . Biochemistry 22 : 4089 - 4095 . (10.1021/bi00286a015) / Biochemistry / Nucleotide sequence of a gene from the Pseudomonas transposon Tn 501 by Brown N. L. (1983)
  5. Brown , N. L. , T. K Misra , J. N. Wimnie , A. Schmidt , M. Seiff , and S. Silver . 1986 . The nucleotide sequence of the mercuric resistance operons of plasmid R100 and transposon TnSOl: further evidence for mer genes which enhance the activity of the mercuric ion detoxification system . Mol. Gen. Genet. 202 : 143 - 151 . (10.1007/BF00330531) / Mol. Gen. Genet. / The nucleotide sequence of the mercuric resistance operons of plasmid R100 and transposon TnSOl: further evidence for mer genes which enhance the activity of the mercuric ion detoxification system by Brown N. L. (1986)
  6. Chang , A. C. Y. , and S. Cohen . 1978 . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P1SA cryptic miniplasmid . J. Bacteriol. 134 : 1141 - 1156 . (10.1128/JB.134.3.1141-1156.1978) / J. Bacteriol. / Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P1SA cryptic miniplasmid by Chang A. C. Y. (1978)
  7. Dretzen , G. , M. Bellard , P. Sassone-Corsi , and P. Chambon . 1981 . A reliable method for the recovery of DNA fragments from agarose and acrylamide gels . Anal. Biochem. 112 : 295 - 298 . (10.1016/0003-2697(81)90296-7) / Anal. Biochem. / A reliable method for the recovery of DNA fragments from agarose and acrylamide gels by Dretzen G. (1981)
  8. Foster , T. J. 1977 . Insertion of the tetracycline resistance translocation unit TnlO in the lac operon of Escherichia coli K12 . Mol. Gen. Genet. 154 : 305 - 309 . (10.1007/BF00571287) / Mol. Gen. Genet. / Insertion of the tetracycline resistance translocation unit TnlO in the lac operon of Escherichia coli K12 by Foster T. J. (1977)
  9. Foster , T. J. , and N. L. Brown . 1985 . Identification of the merR gene of R100 by using mer-lac gene and operon fusions . J. Bacteriol. 163 : 1152 - 1157 . (10.1128/JB.163.3.1153-1157.1985) / J. Bacteriol. / Identification of the merR gene of R100 by using mer-lac gene and operon fusions by Foster T. J. (1985)
  10. Foster , T. J. , and F. Ginnity . 1985 . Some mercurial resistance plasmids from different incompatibility groups specify merR regulatory functions that both repress and induce the mer operon of plasmid R100 . J. Bacteriol. 162 : 773 - 776 . (10.1128/JB.162.2.773-776.1985) / J. Bacteriol. / Some mercurial resistance plasmids from different incompatibility groups specify merR regulatory functions that both repress and induce the mer operon of plasmid R100 by Foster T. J. (1985)
  11. Foster , T. J. , H. Nakahara , A. A. Weiss , and S. Silver . 1979 . Transposon A-generated mutations in the mercuric resistance genes of plasmid R100-1 . J. Bacteriol. 140 : 167 - 181 . (10.1128/JB.140.1.167-181.1979) / J. Bacteriol. / Transposon A-generated mutations in the mercuric resistance genes of plasmid R100-1 by Foster T. J. (1979)
  12. Foster , T. J. , and N. S. Willetts . 1977 . Characterization of transfer-deficient mutants of the R100-1 Tcs plasmid pDU202, caused by insertion of TnlO . Mol. Gen. Genet. 156 : 107 - 114 . (10.1007/BF00272259) / Mol. Gen. Genet. / Characterization of transfer-deficient mutants of the R100-1 Tcs plasmid pDU202, caused by insertion of TnlO by Foster T. J. (1977)
  13. .Gambill B. D. Personal communication.
  14. Gambill , B. D. , and A. 0. Summers . 1992 . Synthesis and degradation of the mRNA of the Tn 2l mer operon . J. Mol. Biol. 225 : 251 - 259 . (10.1016/0022-2836(92)90919-B) / J. Mol. Biol. / Synthesis and degradation of the mRNA of the Tn 2l mer operon by Gambill B. D. (1992)
  15. Gilbert , M. P. , and A. 0. Summers . 1988 . The distribution and divergence of DNA sequences related to the Tn 2l and TnSOl operons . Plasmid 20 : 127 - 136 . (10.1016/0147-619X(88)90015-7) / Plasmid / The distribution and divergence of DNA sequences related to the Tn 2l and TnSOl operons by Gilbert M. P. (1988)
  16. Heltzel , A. , D. Gambili , W. J. Jackson , P. A. Totis , and A. 0. Summers . 1987 . Overexpression and DNA-binding properties of the mer-encoded regulatory protein from plasmid NR1 (Tn 2l) . J. Bacteriol. 169 : 3379 - 3384 . (10.1128/JB.169.7.3379-3384.1987) / J. Bacteriol. / Overexpression and DNA-binding properties of the mer-encoded regulatory protein from plasmid NR1 (Tn 2l) by Heltzel A. (1987)
  17. Inoue , C. , K. Sugawara , and T. Kusano . 1991 . The merR regulatory gene in Thiobacillus ferrooxidans is spaced far apart from the mer structural genes . Mol. Microbiol. 5 : 2707 - 2718 . (10.1111/j.1365-2958.1991.tb01979.x) / Mol. Microbiol. / The merR regulatory gene in Thiobacillus ferrooxidans is spaced far apart from the mer structural genes by Inoue C. (1991)
  18. Jackson , W. J. , and A. 0. Summers . 1982 . Biochemical characterization of HgCl 2-inducible polypeptides encoded by the mer operon of plasmid R100 . J. Bacteriol. 151 : 962 - 970 . (10.1128/JB.151.2.962-970.1982) / J. Bacteriol. / Biochemical characterization of HgCl 2-inducible polypeptides encoded by the mer operon of plasmid R100 by Jackson W. J. (1982)
  19. Jovin , T. M. 1973 . Multiphasic zone electrophoresis. I. Steadystate moving boundary systems formed by different electrolyte combinations . Biochemistry 12 : 871 - 879 . (10.1021/bi00729a014) / Biochemistry / Multiphasic zone electrophoresis. I. Steadystate moving boundary systems formed by different electrolyte combinations by Jovin T. M. (1973)
  20. Jovin , T. M. 1973 . Multiphasic zone electrophoresis. II. Design of integrated discontinuous buffer systems for analytical and preparative fractionation . Biochemistry 12 : 879 - 890 . (10.1021/bi00729a015) / Biochemistry / Multiphasic zone electrophoresis. II. Design of integrated discontinuous buffer systems for analytical and preparative fractionation by Jovin T. M. (1973)
  21. Jovin , T. M. 1973 . Multiphasic zone electrophoresis. III. Further analysis and new forms of discontinuous buffer systems . Biochemistry 12 : 890 - 898 . (10.1021/bi00729a016) / Biochemistry / Multiphasic zone electrophoresis. III. Further analysis and new forms of discontinuous buffer systems by Jovin T. M. (1973)
  22. Jovin T. M. M. L. Dante and A. Chrambach. 1970. Multiphasic buffer systems output PB no. 196085 to 196092 and 203016. National Technical Information Service Springfield Va.
  23. Koop , A. H. , M. E. Hartley , and S. Bourgeois . 1987 . A low-copynumber vector utilizing 3-galactosidase for the analysis of gene control elements . Gene 52 : 245 - 256 . (10.1016/0378-1119(87)90051-5) / Gene / A low-copynumber vector utilizing 3-galactosidase for the analysis of gene control elements by Koop A. H. (1987)
  24. Kusano , T. , G. Ji , C. Inoue , and S. Silver . 1990 . Constitutive synthesis of a transport function encoded by the Thiobacillus ferrooxidans merC gene cloned in Escherichia coli . J. Bacteriol. 172 : 2688 - 2962 . (10.1128/JB.172.5.2688-2692.1990) / J. Bacteriol. / Constitutive synthesis of a transport function encoded by the Thiobacillus ferrooxidans merC gene cloned in Escherichia coli by Kusano T. (1990)
  25. .Landale E. C. C. S. Jones N. V. Hamlett and A. 0. Summers. Unpublished data.
  26. Lee , I. W. , B. D. Gambill , and A. 0. Summers . 1989 . Translation of merD in Tn 2l . J. Bacteriol. 171 : 2222 - 2225 . (10.1128/jb.171.4.2222-2225.1989) / J. Bacteriol. / Translation of merD in Tn 2l by Lee I. W. (1989)
  27. Lund , P. A. , and N. L. Brown . 1987 . Role of the merT and merP gene products of transposon TnSOI in the induction and expression of resistance to mercuric ions . Gene 52 : 207 - 214 . (10.1016/0378-1119(87)90047-3) / Gene / Role of the merT and merP gene products of transposon TnSOI in the induction and expression of resistance to mercuric ions by Lund P. A. (1987)
  28. Lund , P. A. , S. J. Ford , and N. L. Brown . 1986 . Transcriptional regulation of the mercury-resistance genes of transposon TnSOI . J. Gen. Microbiol. 132 : 465 - 480 . / J. Gen. Microbiol. / Transcriptional regulation of the mercury-resistance genes of transposon TnSOI by Lund P. A. (1986)
  29. Maniatis T. E. F. Fritsch and J. Sambrook. 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  30. Merril , C. R. , D. Goldman , S. A. Sedman , and M. H. Ebert . 1981 . Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins . Science 211 : 1437 - 1438 . (10.1126/science.6162199) / Science / Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins by Merril C. R. (1981)
  31. Messing , J. , R. Crea , and P. H. Seeburg . 1981 . A system for shotgun DNA sequencing . Nucleic Acids Res. 9 : 309 - 321 . (10.1093/nar/9.2.309) / Nucleic Acids Res. / A system for shotgun DNA sequencing by Messing J. (1981)
  32. Miller J. H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  33. Misra , T. K. 1992 . Bacterial resistance to inorganic mercury salts and organomercurials . Plasmid 27 : 4 - 16 . (10.1016/0147-619X(92)90002-R) / Plasmid / Bacterial resistance to inorganic mercury salts and organomercurials by Misra T. K. (1992)
  34. Misra , T. K. , N. L. Brown , D. C. Fritzinger , R. D. Pridmore , W. M. Barnes , L. Haberstroh , and S. Silver . 1984 . Mercuric ion-resistance operons of plasmid R100 and transposon TnSOJ: the beginning of the operon including the regulatory region and the first two structural genes . Proc. Natl. Acad. Sci. USA 81 : 5975 - 5979 . (10.1073/pnas.81.19.5975) / Proc. Natl. Acad. Sci. USA / Mercuric ion-resistance operons of plasmid R100 and transposon TnSOJ: the beginning of the operon including the regulatory region and the first two structural genes by Misra T. K. (1984)
  35. Misra , T. K. , N. L. Brown , L. Haberstroh , A. Schmidt , D. Godette , and S. Silver . 1985 . Sequence of the mercuric reductase structural genes from plasmid R100 and transposon TnSOI: functional domains of the enzyme . Gene 34 : 253 - 262 . (10.1016/0378-1119(85)90134-9) / Gene / Sequence of the mercuric reductase structural genes from plasmid R100 and transposon TnSOI: functional domains of the enzyme by Misra T. K. (1985)
  36. Mukhopadhyay , D. H. , H. Yu , G. Nucifora , and T. K Misra . 1991 . Purification and functional characterization of MerD: a coregulator of the mercury resistance operon in Gram-negative bacteria . J. Biol. Chem. 266 : 18538 - 18542 . (10.1016/S0021-9258(18)55095-X) / J. Biol. Chem. / Purification and functional characterization of MerD: a coregulator of the mercury resistance operon in Gram-negative bacteria by Mukhopadhyay D. H. (1991)
  37. Nakahara , H. , S. Silver , T. Mild , and R. H. Rownd . 1979 . Hypersensitivity to Hg 2e and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of the plasmid NR1 . J. Bacteriol. 140 : 161 - 166 . (10.1128/JB.140.1.161-166.1979) / J. Bacteriol. / Hypersensitivity to Hg 2e and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of the plasmid NR1 by Nakahara H. (1979)
  38. Ni'Bhriain , N. , and T. J. Foster . 1986 . Polypeptides specified by the mercuric resistance (mer) operon of plasmid R100 . Gene 32 : 323 - 330 . (10.1016/0378-1119(86)90236-2) / Gene / Polypeptides specified by the mercuric resistance (mer) operon of plasmid R100 by Ni'Bhriain N. (1986)
  39. Ni'Bhriain , N. N. , S. Silver , and T. J. Foster . 1983 . TnS insertion mutations in the mercuric ion resistance genes derived from plasmid R100 . J. Bacteriol. 155 : 690 - 703 . (10.1128/JB.155.2.690-703.1983) / J. Bacteriol. / TnS insertion mutations in the mercuric ion resistance genes derived from plasmid R100 by Ni'Bhriain N. N. (1983)
  40. Novick , R. P. , R. C. Clowes , S. N. Cohen , R. Curtiss m, N. Datta , and S. Falkow . 1976 . Uniform nomenclature of bacterial plasmids: a proposal . Bacteriol. Rev. 40 : 168 - 189 . (10.1128/br.40.1.168-189.1976) / Bacteriol. Rev. / Uniform nomenclature of bacterial plasmids: a proposal by Novick R. P. (1976)
  41. Nucifora , G. , S. Silver , and T. K. Misra . 1990 . Down regulation of the mercury resistance operon by the most promoter-distal gene merD . Mol. Gen. Genet. 220 : 69 - 72 . (10.1007/BF00260858) / Mol. Gen. Genet. / Down regulation of the mercury resistance operon by the most promoter-distal gene merD by Nucifora G. (1990)
  42. O'Halloran , T. V. , and C. Walsh . 1987 . Metalloregulatory DNA-binding protein encoded by the merR gene: isolation and characterization . Science 235 : 211 - 214 . (10.1126/science.3798107) / Science / Metalloregulatory DNA-binding protein encoded by the merR gene: isolation and characterization by O'Halloran T. V. (1987)
  43. Olson , B. H. , S. M. Cayless , S. Ford , and J. N. Lester . 1991 . Toxic element contamination and the occurrence of mercury- resistant bacteria in Hg-contaminated soil, sediments, and sludges . Arch. Environ. Contam. Toxicol. 20 : 226 - 233 . (10.1007/BF01055908) / Arch. Environ. Contam. Toxicol. / Toxic element contamination and the occurrence of mercury- resistant bacteria in Hg-contaminated soil, sediments, and sludges by Olson B. H. (1991)
  44. Olson , B. H. , J. N. Lester , S. M. Cayless , and S. Ford . 1989 . Distribution of mercury resistance determinants in bacterial communities of river sediments . Water Res. 23 : 1209 - 1217 . (10.1016/0043-1354(89)90183-8) / Water Res. / Distribution of mercury resistance determinants in bacterial communities of river sediments by Olson B. H. (1989)
  45. Rickenberg , H. V. , G. N. Cohen , G. Buttin , and J. Monod . 1956 . La galactoside-permease d'Eschenchia coli . Ann. Inst. Pasteur. 91 : 829 - 857 . / Ann. Inst. Pasteur. / La galactoside-permease d'Eschenchia coli by Rickenberg H. V. (1956)
  46. Silver , S. , and M. Walderhaug . 1992 . Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria . Microbiol. Rev. 56 : 195 - 228 . (10.1128/MR.56.1.195-228.1992) / Microbiol. Rev. / Gene regulation of plasmid- and chromosome-determined inorganic ion transport in bacteria by Silver S. (1992)
  47. Struhl , K. 1985 . A rapid method for creating recombinant DNA molecules . BioTechniques 3 : 452 - 453 . / BioTechniques / A rapid method for creating recombinant DNA molecules by Struhl K. (1985)
  48. Summers , A. 0. 1986 . Organization, expression, and evolution of genes for mercury resistance. Annu . Rev. Microbiol. 40 : 607 - 634 . (10.1146/annurev.mi.40.100186.003135) / Rev. Microbiol. / Organization, expression, and evolution of genes for mercury resistance. Annu by Summers A. (1986)
  49. Summers , A. 0. 1992 . Untwist and shout: a heavy metalresponsive transcriptional regulator . J. Bacteriol. 174 : 3097 - 3101 . (10.1128/jb.174.10.3097-3101.1992) / J. Bacteriol. / Untwist and shout: a heavy metalresponsive transcriptional regulator by Summers A. (1992)
  50. Summers , A. O. , and L. Kight-Olliff . 1980 . Tnl generated mutants in the mercuric ion reductase of the Inc P plasmid, R702 . Mol. Gen. Genet. 180 : 91 - 97 . (10.1007/BF00267356) / Mol. Gen. Genet. / Tnl generated mutants in the mercuric ion reductase of the Inc P plasmid, R702 by Summers A. O. (1980)
  51. Summers , A. O. , L. Kight-Olliff , and C. Slater . 1982 . Effect of catabolite repression on the mer operon . J. Bacteriol. 149 : 191 - 197 . (10.1128/JB.149.1.191-197.1982) / J. Bacteriol. / Effect of catabolite repression on the mer operon by Summers A. O. (1982)
Dates
Type When
Created 8 years, 9 months ago (Nov. 7, 2016, 11:07 a.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:15 p.m.)
Indexed 6 days, 17 hours ago (Aug. 31, 2025, 6:35 a.m.)
Issued 32 years, 11 months ago (Oct. 1, 1992)
Published 32 years, 11 months ago (Oct. 1, 1992)
Published Print 32 years, 11 months ago (Oct. 1, 1992)
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@article{Hamlett_1992, title={Roles of the Tn21 merT, merP, and merC gene products in mercury resistance and mercury binding}, volume={174}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.174.20.6377-6385.1992}, DOI={10.1128/jb.174.20.6377-6385.1992}, number={20}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Hamlett, N V and Landale, E C and Davis, B H and Summers, A O}, year={1992}, month=oct, pages={6377–6385} }