Home LiteratureArticle Details
PMID: 2211518 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Identification and cloning of a hemin storage locus involved in the pigmentation phenotype of Yersinia pestis.

Journal of bacteriology ·Vol. 172 ·No. 10 ·1990-10-00 ·Pages 5929-37

Perry RD, Pendrak ML, Schuetze P

Abstract

The temperature-dependent absorption of sufficient exogenous hemin or Congo red to form pigmented colonies of Yersinia pestis has been termed the pigmentation phenotype (Pgm+). Spontaneous mutation to a Pgm- phenotype results in the loss of a number of divergent physiological characteristics, including the ability to store hemin and to bind Congo red at 26 degrees C. In this study, we generated and isolated transposon insertion mutants that are hemin storage negative (Hms-) and therefore unable to form pigmented colonies. These mutations are due to single mini-kan insertions within a 19.5-kilobase (kb) SalI fragment of chromosomal DNA. Restriction site analysis of eight mutants identified a minimum of six potentially different insertion sites spanning an approximately 10-kb hemin storage (hms) locus. The 19.5-kb SalI fragment (containing approximately 18 kb of Y. pestis DNA and the mini-kan insert) was cloned from one of these mutants, KIM6-2012. By using this cloned fragment as a DNA probe, the mechanism of spontaneous mutation to a Pgm- phenotype was identified as a massive deletion event. The deletion spans at least 18 kb of genomic DNA in spontaneous Pgm- mutants from nine separate strains of Y. pestis. DNA adjacent to the mini-kan insert was used to identify a clone containing a wild-type hms locus. A spontaneous Pgm- mutant of Y pestis KIM containing this clone exhibits an Hms+ phenotype. The hms::mini-kan mutations and cloned wild-type hms locus generated in this study will greatly aid in identifying the function of hemin storage in Y. pestis.

MeSH Terms
Biological Transport Blotting, Southern Cloning, Molecular/methods Congo Red/metabolism DNA, Bacterial/genetics,isolation & purification Genes, Bacterial Genotype Heme/metabolism Mutation Phenotype Plasmids Restriction Mapping Temperature Yersinia pestis/genetics,metabolism,physiology
Chemicals
DNA, Bacterial Congo Red Heme
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Perry R D
Department of Microbiology and Immunology, Louisiana State University Medical Center, Shreveport 71130-3932.
Pendrak M L
Schuetze P
References (44)
44 references, click to expand
  1. Cytoplasmic and membrane proteins of yersiniae cultivated under conditions simulating mammalian intracellular environment.
    Proc Natl Acad Sci U S A. 1981 Feb;78(2):1224-8 PMID: 6940138
  2. Essential virulence determinants of different Yersinia species are carried on a common plasmid.
    Plasmid. 1981 Mar;5(2):183-7 PMID: 7243971
  3. Loss of pigmentation in Shigella flexneri 2a is correlated with loss of virulence and virulence-associated plasmid.
    Infect Immun. 1984 Jan;43(1):397-401 PMID: 6360906
  4. Localization in Yersinia pestis of peptides associated with virulence.
    Infect Immun. 1982 Apr;36(1):129-35 PMID: 6210636
  5. A small cosmid for efficient cloning of large DNA fragments.
    Gene. 1980 Nov;11(3-4):291-8 PMID: 6260575
  6. A simple method for the preparation of large quantities of pure plasmid DNA.
    Biochim Biophys Acta. 1975 Apr 2;383(4):457-63 PMID: 1092355
  7. Genetic analysis of the 9.5-kilobase virulence plasmid of Yersinia pestis.
    Infect Immun. 1988 Oct;56(10):2743-8 PMID: 2843470
  8. In vivo repackaging of recombinant cosmid molecules for analyses of Salmonella typhimurium, Streptococcus mutans, and mycobacterial genomic libraries.
    Infect Immun. 1986 Apr;52(1):101-9 PMID: 2937735
  9. Specificity, induction, and absorption of pesticin.
    J Bacteriol. 1972 Oct;112(1):212-9 PMID: 4627922
  10. In vivo comparison of avirulent Vwa- and Pgm- or Pstr phenotypes of yersiniae.
    Infect Immun. 1984 Mar;43(3):895-900 PMID: 6365786
  11. Outer membrane peptides of Yersinia pestis mediating siderophore-independent assimilation of iron.
    Biol Met. 1989;2(3):174-84 PMID: 2535180
  12. Molecular alteration of the 140-megadalton plasmid associated with loss of virulence and Congo red binding activity in Shigella flexneri.
    Infect Immun. 1986 Feb;51(2):470-5 PMID: 3002985
  13. The Vwa+ virulence factor of yersiniae: the molecular basis of the attendant nutritional requirement for Ca++.
    Rev Infect Dis. 1983 Sep-Oct;5 Suppl 4:S748-58 PMID: 6195719
  14. Versatile low-copy-number plasmid vectors for cloning in Escherichia coli.
    Gene. 1982 Jun;18(3):335-41 PMID: 6290337
  15. High efficiency transformation of E. coli by high voltage electroporation.
    Nucleic Acids Res. 1988 Jul 11;16(13):6127-45 PMID: 3041370
  16. Pesticins. 3. Expression of coagulase and mechanism of fibrinolysis.
    J Bacteriol. 1967 Jul;94(1):19-26 PMID: 6027989
  17. Congo red-mediated regulation of levels of Shigella flexneri 2a membrane proteins.
    Infect Immun. 1989 Aug;57(8):2364-71 PMID: 2663721
  18. Molecular cloning of a genetic determinant for Congo red binding ability which is essential for the virulence of Shigella flexneri.
    Infect Immun. 1986 Feb;51(2):476-82 PMID: 3510977
  19. Accumulation of iron by yersiniae.
    J Bacteriol. 1979 Mar;137(3):1290-8 PMID: 108243
  20. The gene coding for the 190,000-dalton iron-regulated protein of Yersinia species is present only in the highly pathogenic strains.
    Infect Immun. 1989 Apr;57(4):1211-7 PMID: 2466794
  21. The virulence-enhancing effect of iron on nonpigmented mutants of virulent strains of Pasteurella pestis.
    Br J Exp Pathol. 1956 Dec;37(6):577-83 PMID: 13396142
  22. Pesticins. I. Pesticinbacterium interrelationships, and environmental factors influencing activity.
    J Bacteriol. 1961 Dec;82:940-9 PMID: 13873808
  23. Genetic analysis of the low calcium response in Yersinia pestis mu d1(Ap lac) insertion mutants.
    J Bacteriol. 1984 Dec;160(3):842-8 PMID: 6094509
  24. Rapid procedure for detection and isolation of large and small plasmids.
    J Bacteriol. 1981 Mar;145(3):1365-73 PMID: 7009583
  25. Detection and differentiation of iron-responsive avirulent mutants on Congo red agar.
    Infect Immun. 1977 Oct;18(1):94-8 PMID: 409688
  26. Differential clearance and host-pathogen interactions of YopE- and YopK- YopL- Yersinia pestis in BALB/c mice.
    Infect Immun. 1989 Apr;57(4):1200-10 PMID: 2925246
  27. Mutation rate to nonpigmentation in Pasteurella pestis.
    J Bacteriol. 1969 Jun;98(3):1404-6 PMID: 5788712
  28. Resistance to pesticin, storage of iron, and invasion of HeLa cells by Yersiniae.
    Infect Immun. 1987 Mar;55(3):572-8 PMID: 3818085
  29. Congo red-agar plating medium for detecting pigmentation in Pasteurella pestis.
    Appl Microbiol. 1969 Nov;18(5):834-7 PMID: 5370459
  30. A rapid alkaline extraction procedure for screening recombinant plasmid DNA.
    Nucleic Acids Res. 1979 Nov 24;7(6):1513-23 PMID: 388356
  31. Congo red medium to distinguish between invasive and non-invasive Escherichia coli pathogenic for poultry.
    Avian Dis. 1986 Jan-Mar;30(1):117-21 PMID: 3524540
  32. A 101-kilodalton heme-binding protein associated with congo red binding and virulence of Shigella flexneri and enteroinvasive Escherichia coli strains.
    Infect Immun. 1989 Nov;57(11):3534-9 PMID: 2680975
  33. Vwa+ phenotype of Yersinia enterocolitica.
    Infect Immun. 1983 Apr;40(1):166-71 PMID: 6832830
  34. Differentiation between virulent and avirulent Yersinia enterocolitica isolates by using Congo red agar.
    J Clin Microbiol. 1983 Sep;18(3):486-90 PMID: 6630438
  35. Phase variants of Bordetella bronchiseptica arise by spontaneous deletions in the vir locus.
    Mol Microbiol. 1989 Dec;3(12):1719-28 PMID: 2560120
  36. Plasmids in Yersinia pestis.
    Infect Immun. 1981 Feb;31(2):839-41 PMID: 7216478
  37. Plasmids of Escherichia coli as cloning vectors.
    Methods Enzymol. 1979;68:245-67 PMID: 232214
  38. New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition.
    Gene. 1984 Dec;32(3):369-79 PMID: 6099322
  39. Congo red binding phenotype is associated with hemin binding and increased infectivity of Shigella flexneri in the HeLa cell model.
    Infect Immun. 1987 Jun;55(6):1393-8 PMID: 3553000
  40. Virulence genes regulated at the transcriptional level by Ca2+ in Yersinia pestis include structural genes for outer membrane proteins.
    Infect Immun. 1986 Feb;51(2):445-54 PMID: 3002984
  41. The pigmentation of Pasteurella pestis on a defined medium containing haemin.
    Br J Exp Pathol. 1956 Dec;37(6):570-6 PMID: 13396141
  42. Mutations to tolerance and resistance to pesticin and colicins in Escherichia coli phi.
    J Bacteriol. 1981 May;146(2):506-11 PMID: 7012131
  43. Deletion of a 55-kilobase-pair DNA element from the chromosome during heterocyst differentiation of Anabaena sp. strain PCC 7120.
    J Bacteriol. 1988 Nov;170(11):5034-41 PMID: 3141375
  44. A low-Ca2+ response operon encodes the V antigen of Yersinia pestis.
    Infect Immun. 1986 Nov;54(2):428-34 PMID: 3021629
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.172.10.5929-5937.1990
Published
1990-10-00
Pages
5929-37
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC526914
Subset
IM
Grants
NIAID NIH HHS · AI25098 · United States
External Links
PubMed source DOI: 10.1128/jb.172.10.5929-5937.1990 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2