Structure and regulation of the Yersinia pestis yscBCDEF operon
10.1128/jb.174.14.4820-4828.1992
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

We have investigated the physical and genetic structure and regulation of the Yersinia pestis yscBCDEF region, previously called lcrC. DNA sequence analysis showed that this region is homologous to the corresponding part of the ysc locus of Yersinia enterocolitica and suggested that the yscBCDEF cistrons belong to a single operon on the low-calcium response virulence plasmid pCD1. Promoter activity measurements of ysc subclones indicated that yscBCDEF constitutes a suboperon of the larger ysc region by revealing promoter activity in a clone containing the 3' end of yscD, intact yscE and yscF, and part of yscG. These experiments also revealed an additional weak promoter upstream of yscD. Northern (RNA) analysis with a yscD probe showed that operon transcription is thermally induced and downregulated in the presence of Ca2+. Primer extension of operon transcripts suggested that two promoters, a moderate-level constitutive one and a stronger, calcium-downregulated one, control full-length operon transcription at 37 degrees C. Primer extension provided additional support for the proposed designation of a yscBCDEF suboperon by identifying a 5' end within yscF, for which relative abundances in the presence and absence of Ca2+ revealed regulation that is distinct from that for transcripts initiating farther upstream. YscB and YscC were expressed in Escherichia coli by using a high-level transcription system. Attempts to express YscD were only partially successful, but they revealed interesting regulation at the translational level.

Bibliography

Haddix, P. L., & Straley, S. C. (1992). Structure and regulation of the Yersinia pestis yscBCDEF operon. Journal of Bacteriology, 174(14), 4820–4828.

Authors 2
  1. P L Haddix (first)
  2. S C Straley (additional)
References 53 Referenced 48
  1. Bergman , T. , S. H Akansson , A. Forsberg , L. Norlander , A. Macellaro , A. Backman , I. Bolin , and H. Wolf-Watz . 1991 . Analysis of the V antigen IcrGVH-yopBD operon of Yersinia pseudotuberculosis: evidence for a regulatory role of LcrH and LcrV . J. Bacteriol. 173 : 1607 - 1616 . (10.1128/JB.173.5.1607-1616.1991) / J. Bacteriol. / Analysis of the V antigen IcrGVH-yopBD operon of Yersinia pseudotuberculosis: evidence for a regulatory role of LcrH and LcrV by Bergman T. (1991)
  2. Birnboim , H. C. , and J. Doly . 1979 . A rapid alkaline extraction procedure for screening recombinant plasmid DNA . Nucleic Acids Res. 7 : 1513 - 1523 . (10.1093/nar/7.6.1513) / Nucleic Acids Res. / A rapid alkaline extraction procedure for screening recombinant plasmid DNA by Birnboim H. C. (1979)
  3. Bliska , J. B. , K. Guan , J. E. Dixon , and S. Falkow . 1991 . Tyrosine phosphate hydrolysis of host proteins by an essential Yersinia virulence determinant . Proc. Natl. Acad. Sci. USA 88 : 1187 - 1191 . (10.1073/pnas.88.4.1187) / Proc. Natl. Acad. Sci. USA / Tyrosine phosphate hydrolysis of host proteins by an essential Yersinia virulence determinant by Bliska J. B. (1991)
  4. Bolivar , F. , and K. Backman . 1979 . Plasmids of Eschenchia coli as cloning vectors . Methods Enzymol. 68 : 245 - 267 . (10.1016/0076-6879(79)68018-7) / Methods Enzymol. / Plasmids of Eschenchia coli as cloning vectors by Bolivar F. (1979)
  5. Bolivar , F. , R. L. Rodriguez , P. J. Greene , M. C. Betlach , H. L. Heynecker , H. W. Boyer , J. H. Crosa , and S. Falkow . 1977 . Construction and characterization of new cloning vehicles. II. A multipurpose cloning system . Gene 2 : 95 - 113 . (10.1016/0378-1119(77)90000-2) / Gene / Construction and characterization of new cloning vehicles. II. A multipurpose cloning system by Bolivar F. (1977)
  6. Brubaker R. R. 1986. Low-calcium response of virulent yersiniae p. 43-48. In L. Leive P. F. Bonventre J. A. Morello S. D. Silver and H. C. Wu (ed.) Microbiology-1986. American Society for Microbiology Washington D.C.
  7. Castilho , B. A. , P. Olfson , and M. J. Casadaban . 1984 . Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons . J. Bacteriol. 158 : 488 - 495 . (10.1128/jb.158.2.488-495.1984) / J. Bacteriol. / Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons by Castilho B. A. (1984)
  8. Chung , C. T. , S. L. Niemela , and R. H. Miller . 1989 . One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution . Proc. Natl. Acad. Sci. USA 86 : 2172 - 2175 . (10.1073/pnas.86.7.2172) / Proc. Natl. Acad. Sci. USA / One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution by Chung C. T. (1989)
  9. Cornelis , G. , Y. Laroche , G. Balligand , M. -P. Sory , and G. Wauters . 1987 . Yersinia enterocolitica, a primary model for bacterial invasiveness . Rev. Infect. Dis. 9 : 64 - 87 . (10.1093/clinids/9.1.64) / Rev. Infect. Dis. / Yersinia enterocolitica, a primary model for bacterial invasiveness by Cornelis G. (1987)
  10. Cornelis , G. , C. Sluiters , C. L. de Rouvroit , and T. Michiels . 1989 . Homology between VirF, the transcriptional activator of the Yersinia virulence regulon, and AraC, the Escherichia coli arabinose operon regulator . J. Bacteriol. 171 : 254 - 262 . (10.1128/jb.171.1.254-262.1989) / J. Bacteriol. / Homology between VirF, the transcriptional activator of the Yersinia virulence regulon, and AraC, the Escherichia coli arabinose operon regulator by Cornelis G. (1989)
  11. Cornelis , G. , M. -P. Sory , Y. Laroche , and I. Derclaye . 1986 . Genetic analysis of the plasmid region controlling virulence in Yersinia enterocolitica 0:9 by mini-mu insertions and lac gene fusions . Microb. Pathog. 1 : 349 - 359 . (10.1016/0882-4010(86)90067-7) / Microb. Pathog. / Genetic analysis of the plasmid region controlling virulence in Yersinia enterocolitica 0:9 by mini-mu insertions and lac gene fusions by Cornelis G. (1986)
  12. Cornelis , G. R. , T. Biot , C. Lambert de Rouvroit , T. Michiels , B. Mulder , C. Sluiters , M. -P. Sory , M. Van Bouchaute , and J. -C. Vanooteghem . 1991 . The Yersinia yop regulon . Mol. Microbiol. 3 : 1455 - 1459 . (10.1111/j.1365-2958.1989.tb00129.x) / Mol. Microbiol. / The Yersinia yop regulon by Cornelis G. R. (1991)
  13. Davis R. H. D. Botstein and J. R. Roth. 1980. Advanced bacterial genetics. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  14. Goguen , J. D. , J. Yother , and S. C. Straley . 1984 . Genetic analysis of the low calcium response in Yersinia pestis Mudl (Ap lac) insertion mutants . J. Bacteriol. 160 : 842 - 848 . (10.1128/JB.160.3.842-848.1984) / J. Bacteriol. / Genetic analysis of the low calcium response in Yersinia pestis Mudl (Ap lac) insertion mutants by Goguen J. D. (1984)
  15. Gold L. and G. Stormo. 1987. Translational initiation p. 1302-1307. In F. C. Neidhardt J. L. Ingraham K. B. Low B. Magasanik M. Schaechter and H. E. Umbarger (ed.) Escherichia coli and Salmonella typhimurium: cellular and molecular biology vol. 2. American Society for Microbiology Washington D.C.
  16. Guan , K. , and J. E. Dixon . 1990 . Protein tyrosine phosphatase activity of an essential virulence determinant in Yersinia . Science 249 : 553 - 556 . (10.1126/science.2166336) / Science / Protein tyrosine phosphatase activity of an essential virulence determinant in Yersinia by Guan K. (1990)
  17. Haddix P. L. and S. C. Straley. The Yersinia pestis yscD gene encodes a membrane protein which functions in secretion and regulation in the low Ca 2' response. Submitted for publication.
  18. . Hoe , N. P. , C. F. Minion , and J. D. Goguen . 1992 . Temperature sensing in Yersinia pestis: regulation of yopE transcription by 1crF . J. Bacteriol. 174 : 4275 - 4286 . (10.1128/JB.174.13.4275-4286.1992) / J. Bacteriol. / Temperature sensing in Yersinia pestis: regulation of yopE transcription by 1crF by Hoe N. P. (1992)
  19. Kado , C. I. , and S. T. Liu . 1981 . Rapid procedure for detection and isolation of large and small plasmids . J. Bacteriol. 145 : 1365 - 1373 . (10.1128/JB.145.3.1365-1373.1981) / J. Bacteriol. / Rapid procedure for detection and isolation of large and small plasmids by Kado C. I. (1981)
  20. Krueger , J. H. , and G. C. Walker . 1983 . Mud (Ap lac)-generated fusions in studies of gene expression . Methods Enzymol. 100 : 501 - 509 . (10.1016/0076-6879(83)00075-0) / Methods Enzymol. / Mud (Ap lac)-generated fusions in studies of gene expression by Krueger J. H. (1983)
  21. Laemmli , U. K. 1970 . Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature (London) 227 : 680 - 685 . (10.1038/227680a0) / Nature (London) / Cleavage of structural proteins during the assembly of the head of bacteriophage T4 by Laemmli U. K. (1970)
  22. Lawton , W. D. , R. L. Erdman , and M. J. Surgalla . 1963 . Biosynthesis and purification of v and w antigen in Pasteurella pestis . J. Immunol. 91 : 179 - 184 . (10.4049/jimmunol.91.2.179) / J. Immunol. / Biosynthesis and purification of v and w antigen in Pasteurella pestis by Lawton W. D. (1963)
  23. Leung , K. Y. , B. S. Reisner , and S. C. Straley . 1990 . YopM inhibits platelet aggregation and is necessary for virulence of Yersinia pestis in mice . Infect. Immun. 58 : 3262 - 3271 . (10.1128/IAI.58.10.3262-3271.1990) / Infect. Immun. / YopM inhibits platelet aggregation and is necessary for virulence of Yersinia pestis in mice by Leung K. Y. (1990)
  24. Leung , K. Y. , and S. C. Straley . 1989 . The yopM gene of Yersinia pestis encodes a released protein having homology with the human platelet surface protein GPIba . J. Bacteriol. 171 : 4623 - 4632 . (10.1128/jb.171.9.4623-4632.1989) / J. Bacteriol. / The yopM gene of Yersinia pestis encodes a released protein having homology with the human platelet surface protein GPIba by Leung K. Y. (1989)
  25. Lindler , L. E. , M. S. Klempner , and S. C. Straley . 1990 . Yersinia pestis pH6 antigen: genetic, biochemical, and virulence characterization of a protein involved in the pathogenesis of bubonic plague . Infect. Immun. 58 : 2569 - 2577 . (10.1128/IAI.58.8.2569-2577.1990) / Infect. Immun. / Yersinia pestis pH6 antigen: genetic, biochemical, and virulence characterization of a protein involved in the pathogenesis of bubonic plague by Lindler L. E. (1990)
  26. [inn, T., and R. St. Pierre . 1990 . Improved vector system for constructing transcriptional fusions that ensures independent translation of lacZ . J. Bacteriol. 172 : 1077 - 1084 . (10.1128/JB.172.2.1077-1084.1990) / J. Bacteriol. / Improved vector system for constructing transcriptional fusions that ensures independent translation of lacZ by St. Pierre (1990)
  27. Maniatis T. E. F. Fritsch and J. Sambrook 1982. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  28. Michiels , T. , J. -C. Vanooteghem , C. L. de Rouvroit , B. China , A. Gustin , A. Gustin , P. Boudry , and G. R. Cornelis . 1991 . Analysis of virC, an operon involved in the secretion of Yop proteins by Yersinia enterocolitica . J. Bacteriol. 173 : 4994 - 5009 . (10.1128/JB.173.16.4994-5009.1991) / J. Bacteriol. / Analysis of virC, an operon involved in the secretion of Yop proteins by Yersinia enterocolitica by Michiels T. (1991)
  29. Michiels , T. , P. Wattiau , R. Brasseur , J. -M. Ruysschaert , and G. Cornelis . 1990 . Secretion of Yop proteins by yersiniae . Infect. Immun. 58 : 2840 - 2849 . (10.1128/IAI.58.9.2840-2849.1990) / Infect. Immun. / Secretion of Yop proteins by yersiniae by Michiels T. (1990)
  30. Miller J. H. 1972. Experiments in molecular genetics p. 352-359. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  31. Mulder , B. , T. Michiels , M. Simonet , M. -P. Sory , and G. Cornelis . 1989 . Identification of additional virulence determinants on the pYV plasmid of Yersinia enterocolitica W227 . Infect. Immun. 57 : 2534 - 2541 . (10.1128/IAI.57.8.2534-2541.1989) / Infect. Immun. / Identification of additional virulence determinants on the pYV plasmid of Yersinia enterocolitica W227 by Mulder B. (1989)
  32. O'Connor , M. B. , and M. H. Malamy . 1983 . A new insertion sequence, IS121, is found on the MudIl (Ap lac) bacteriophage and the Escherichia coli K-12 chromosome . J. Bacteriol. 156 : 669 - 679 . (10.1128/JB.156.2.669-679.1983) / J. Bacteriol. / A new insertion sequence, IS121, is found on the MudIl (Ap lac) bacteriophage and the Escherichia coli K-12 chromosome by O'Connor M. B. (1983)
  33. Perry , R. D. , P. Haddix , E. B. Atkins , T. K Soughers , and S. C. Straley . 1987 . Regulation of expression of V antigen and outer membrane proteins in Yersinia pestis . Contrib. Microbiol. Immunol. 9 : 173 - 178 . / Contrib. Microbiol. Immunol. / Regulation of expression of V antigen and outer membrane proteins in Yersinia pestis by Perry R. D. (1987)
  34. Perry , R. D. , M. Pendrak , and P. Schuetze . 1990 . Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis . J. Bacteriol. 172 : 5929 - 5937 . (10.1128/JB.172.10.5929-5937.1990) / J. Bacteriol. / Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis by Perry R. D. (1990)
  35. Piano , G. V. , S. S. Barve , and S. C. Straley . 1991 . LcrD, a membrane-bound regulator of the Yersinia pestis low-calcium response . J. Bacteriol. 173 : 7293 - 7303 . (10.1128/JB.173.22.7293-7303.1991) / J. Bacteriol. / LcrD, a membrane-bound regulator of the Yersinia pestis low-calcium response by Piano G. V. (1991)
  36. Portnoy , D. A. , H. F. Blank , D. T. Kingsbury , and S. Falkow . 1983 . Genetic analysis of essential plasmid determinants of pathogenicity in Yersinia pestis . J. Infect. Dis. 148 : 297 - 304 . (10.1093/infdis/148.2.297) / J. Infect. Dis. / Genetic analysis of essential plasmid determinants of pathogenicity in Yersinia pestis by Portnoy D. A. (1983)
  37. Portnoy , D. A. , and R. J. Martinez . 1985 . Role of a plasmid in the pathogenicity of Yersinia species . Curr. Top. Microbiol. Immunol. 118 : 29 - 51 . / Curr. Top. Microbiol. Immunol. / Role of a plasmid in the pathogenicity of Yersinia species by Portnoy D. A. (1985)
  38. Price , S. B. , C. Cowan , R. D. Perry , and S. C. Straley . 1991 . The Yersinia pestis V antigen is a regulatory protein necessary for Ca 2"-dependent growth and maximal expression of low Ca 2l response virulence genes . J. Bacteriol. 173 : 2649 - 2657 . (10.1128/JB.173.8.2649-2657.1991) / J. Bacteriol. / The Yersinia pestis V antigen is a regulatory protein necessary for Ca 2"-dependent growth and maximal expression of low Ca 2l response virulence genes by Price S. B. (1991)
  39. Rosqvist , R. , I. B61in, and H. Wolf-Watz . 1988 . Inhibition of phagocytosis in Yersinia pseudotuberculosis: a virulence plasmid-encoded ability involving the Yop 2b protein . Infect. Immun. 56 : 2139 - 2143 . (10.1128/IAI.56.8.2139-2143.1988) / Infect. Immun. / Inhibition of phagocytosis in Yersinia pseudotuberculosis: a virulence plasmid-encoded ability involving the Yop 2b protein by Rosqvist R. (1988)
  40. Rosqvist , R. , A. Forsberg , M. Rimpilainen , T. Bergman , and H. Wolf-Watz . 1990 . The cytotoxic protein YopE of Yersinia obstructs the primary host defence . Mol. Microbiol. 4 : 657 - 667 . (10.1111/j.1365-2958.1990.tb00635.x) / Mol. Microbiol. / The cytotoxic protein YopE of Yersinia obstructs the primary host defence by Rosqvist R. (1990)
  41. Rosqvist , R. , A. Forsberg , and H. Wolf-Watz . 1991 . Intracellular targetting of the Yersina YopE cytotoxin in mammalian cells induces actin microfilament disruption . Infect. Immun. 59 : 4562 - 4569 . (10.1128/IAI.59.12.4562-4569.1991) / Infect. Immun. / Intracellular targetting of the Yersina YopE cytotoxin in mammalian cells induces actin microfilament disruption by Rosqvist R. (1991)
  42. Sanger , F. , S. Nicklen , and A. R. Coulson . 1977 . DNA sequencing with chain-terminating inhibitors . Proc. Natl. Acad. Sci. USA 74 : 5463 - 5467 . (10.1073/pnas.74.12.5463) / Proc. Natl. Acad. Sci. USA / DNA sequencing with chain-terminating inhibitors by Sanger F. (1977)
  43. Seifert , H. S. , E. Y. Chen , M. So , and F. Heifron . 1986 . Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae . Proc. Natl. Acad. Sci. USA 83 : 735 - 739 . (10.1073/pnas.83.3.735) / Proc. Natl. Acad. Sci. USA / Shuttle mutagenesis: a method of transposon mutagenesis for Saccharomyces cerevisiae by Seifert H. S. (1986)
  44. Shine , J. , and L. Dalgarno . 1974 . The 3' terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites . Proc. Natl. Acad. Sci. USA 71 : 1342 - 1346 . (10.1073/pnas.71.4.1342) / Proc. Natl. Acad. Sci. USA / The 3' terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites by Shine J. (1974)
  45. Silhavy T. J. M. L. Berman and L. W. Enquist. 1984. Experiments with gene fusions p. 113-114. Cold Spring Harbor Laboratory Cold Spring Harbor N.Y.
  46. Straley , S. C. 1991 . The low Ca 2" response virulence regulon of human pathogenic Yersiniae . Microb. Pathog. 10 : 87 - 91 . (10.1016/0882-4010(91)90069-M) / Microb. Pathog. / The low Ca 2" response virulence regulon of human pathogenic Yersiniae by Straley S. C. (1991)
  47. Straley , S. C. , and W. S. Bowmer . 1986 . Virulence genes regulated at the transcriptional level by Ca 2+ in Yersinia pestis include structural genes for outer membrane proteins . Infect. Immun. 51 : 445 - 454 . (10.1128/IAI.51.2.445-454.1986) / Infect. Immun. / Virulence genes regulated at the transcriptional level by Ca 2+ in Yersinia pestis include structural genes for outer membrane proteins by Straley S. C. (1986)
  48. Straley , S. C. , and M. L. Cibull . 1989 . Differential clearance and host-pathogen interactions of YopE- and YopK- YopL- Yersinia pestis in BALB/c mice . Infect. Immun. 57 : 1200 - 1210 . (10.1128/IAI.57.4.1200-1210.1989) / Infect. Immun. / Differential clearance and host-pathogen interactions of YopE- and YopK- YopL- Yersinia pestis in BALB/c mice by Straley S. C. (1989)
  49. Une , T. , and R. R. Brubaker . 1984 . In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae . Infect. Immun. 43 : 895 - 900 . (10.1128/IAI.43.3.895-900.1984) / Infect. Immun. / In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae by Une T. (1984)
  50. Une , T. , and R. R. Brubaker . 1984 . Roles of V antigen in promoting virulence and immunity in yersiniae . J. Immunol. 133 : 2226 - 2230 . (10.4049/jimmunol.133.4.2226) / J. Immunol. / Roles of V antigen in promoting virulence and immunity in yersiniae by Une T. (1984)
  51. Vieira , J. , and J. Messing . 1987 . Production of single-stranded plasmid DNA . Methods Enzymol. 153 : 3 - 11 . (10.1016/0076-6879(87)53044-0) / Methods Enzymol. / Production of single-stranded plasmid DNA by Vieira J. (1987)
  52. Viitanen , A. - M. , P. Toivanen , and M. Skurnik . 1990 . The IcrE gene is part of an operon in the Icr region of Yersinia enterocolitica 0:3 . J. Bacteriol. 172 : 3152 - 3162 . (10.1128/JB.172.6.3152-3162.1990) / J. Bacteriol. / The IcrE gene is part of an operon in the Icr region of Yersinia enterocolitica 0:3 by Viitanen A. (1990)
  53. Yother , J. , T. W. Chamness , and J. D. Goguen . 1986 . Temperature-controlled plasmid regulon associated with low calcium response in Yersinia pestis . J. Bacteriol. 165 : 443 - 447 . (10.1128/JB.165.2.443-447.1986) / J. Bacteriol. / Temperature-controlled plasmid regulon associated with low calcium response in Yersinia pestis by Yother J. (1986)
Dates
Type When
Created 8 years, 9 months ago (Nov. 7, 2016, 12:02 p.m.)
Deposited 2 years ago (Aug. 20, 2023, 4:01 p.m.)
Indexed 1 year, 9 months ago (Oct. 28, 2023, 10:49 a.m.)
Issued 33 years, 1 month ago (July 1, 1992)
Published 33 years, 1 month ago (July 1, 1992)
Published Print 33 years, 1 month ago (July 1, 1992)
Funders 0

None

@article{Haddix_1992, title={Structure and regulation of the Yersinia pestis yscBCDEF operon}, volume={174}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.174.14.4820-4828.1992}, DOI={10.1128/jb.174.14.4820-4828.1992}, number={14}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Haddix, P L and Straley, S C}, year={1992}, month=jul, pages={4820–4828} }