Cloning of the groE operon of the marine bacterium Vibrio harveyi using a lambda vector

• Autorzy: Kuchanny D. , Klein G. , Krzewska J. , Czyz A. , Lipinska B.
• Języki publikacji: EN

Abstrakty

EN

groES and groEL genes encode two co-operating proteins GroES and GroEL, belonging to a class of chaperone proteins highly conserved during evolution. The GroE chaperones are indispensable for the growth of bacteriophage λ in Escherichia coli cells. In order to clone the groEL and groES genes of the marine bacterium Vibrio harveyi, we constructed the V. harveyi genomic library in the λEMBL1 vector, and selected clones which were able to complement mutations in both groE genes of E. coli for bacteriophage λ growth. Using Southern hybridization, in one of these clones we identified a DNA fragment homologous to the E. coli groE region. Analysis of the nucleotide sequence of this fragment showed that the cloned region contained a sequence in 71.7% homologous to the 3' end of the groEL gene of E. coli. This confirmed that the λ clone indeed carries the groE region of V. harveyi. The positive result of our strategy of cloning with the use of the genomic library in λ vector suggests that the same method might be useful in the isolation of the groE homologues from other bacteria. The V. harveyi cloned groE genes did not suppress thermosensitivity of the E. coli groE mutants.

Słowa kluczowe

EN

hybrid clone; lambda vector; hybridization; heat shock; Escherichia coli; mutant strain; Vibrio harveyi; marine bacteria

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1998
• Tom: 45
• Numer: 1
• Opis fizyczny: p.261-270,fig.

Twórcy

autor

Kuchanny D.

• University of Gdansk, Kladki 24, 80-822 Gdansk, Poland

autor

Klein G.

autor

Krzewska J.

autor

Czyz A.

autor

Lipinska B.

Bibliografia

• 1. Hartl, F.U. (1996) Molecular chaperones in cellular protein folding. Nature 381,571-580.
• 2. Fayet, O., Ziegelhoffer, T. & Georgopoulos, C. (1989) The groES and groEL heat shock gene products of Escherichia coli are essential for bacterial growth at all temperatures. J. Bacte- rioL 171, 1379-1385.
• 3. Zeilstra-Ryalls, J., Fayet, O. & Georgopoulos, C. (1991) The universally conserved GroE (Hsp60) chaperonins. Annu Rev. Microbiol 45, 301-325.
• 4. Segal, G. & Ron, E.Z. (19%) Regulation and organization of the groE and dnaK operons in Eubacteria. FEMS Microbiol Lett. 138,1 -10.
• 5. Georgopoulos, C., Ang, D., Liberek, K. & Ży­licz, M. (1990) Properties of the Escherichia coli heat shock proteins and their role in bacte­riophage A growth; in Stress Proteins in Biology and Medicine (Morimoto, R.I., Tissieres, A. & Georgopoulos, C., eds.) pp. 191-221, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
• 6. Yura, T., Nagai, H. & Mori, H. (1993) Regula­tion of the heat-shock response in bacteria. Annu. Rev. Microbiol. 47. 321-350.
• 7. Schumann, W. (1996) Regulation of the heat shock response in Escherichia coli and Bacillus subtilis. J. Biosci. 21, 133-148.
• 8. Schumann, W. (1996) The heat shock stimu- lon of Bacillus subtilis. Brazilian J. Genet. 19, 387-398.
• 9. Klein, G., Walczak, R., Krasnowska. E., Blasz- czak, A. & Lipińska, B. (1995) Characteriza­tion of heat-shock response of the marine bac­terium Vibrio harveyi. MoL Microbiol. 16,801- 811.
• 10. Sambrook, J., Fritsch, F. & Maniatis, T.E. (1989) Molecular Cloning: A Laboratory Man­ual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
• 11. Silhavy, T.J., Berman, L. & Enquist, L. W. (1984) in Experiments with Gene Fusions-, pp. 137-139, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
• 12. Miller, J.H. (1992) A Short Course in Bacterial Genetics. Cold Spring Harbor Press. Cold Spring Harbor, N.Y.
• 13. Rusanganwa, E. & Gupta, R.S. (1993) Cloning and characterization of multiple groEL chap­eron in-encoding genes in Rhizobium melUoti. Gene 126, 67-75.
• 14. Arnau, J., Sorensen, K., Appel, K.F., Vo- gensen, F.K. & Hammer, K. (1996) Analysis of heat shock gene expression in Lactococcus lac­tis MG1363. Microbiol. 142, 1685-1691.
• 15. Falah, M. & Gupta, R. S. (1994) Cloning of the hsp70 (dnaK) genes from Rhizobium meliloti and Pseudomonas cepacia: Phylogenetic analy­ses of mitochondrial origin based on a highly conserved protein sequence. J. BacterioL 176, 7748-7753.
• 16. Brans, A., Loriaux, A., Joris, B. & Dusart, «J. (1996) Cloning and sequencing of the dnaK lo­cus in Streptomyces coelicolor A3(2). DNA Se­quence 6, 179-184.
• 17. Eaton, T., Shearman, C. & Gasson, M. (1993) Cloning and sequence analysis of the dnaK gene region of Lactococcus lactis subsp. lactis. J. Gen. Microbiol. 139, 3253-3264.
• 18. Nimura, K., Yoshikawa, H. & Takahashi, H. (1994) Identification of dnaK multigene fam­ily in Synechococcus sp PCC7942. Biochem. Biophys. Res. Commun. 201, 466-471.
• 19. Segal, G. & Ron, E.Z. (1995) The dnaKJ operon of Agrobacterium tumefaciens: Tran­scriptional analysis and evidence for a new heat shock promoter. J. BacterioL 177, 5952- 5958.
• 20. Zuber, M., Hoover, T.A. & Court, D.L. (1995) Cloning, sequencing and expression of the dnaJ gene of Coxiella burnetii. Gene 152, 99-102.
• 21. Segal, G. & Ron. E.Z. (1993) Heat shock tran­scription of the groESL operon of Agrobacte­rium tumefaciens may involve a hairpin-loop structure. J. BacterioL 175, 3083-3088.
• 22. Lee. W.T., Terlesky, K.C. & Tabita, F.R. (1997) Cloning and charactcrization of two groESL operons of Rhodobacter sphaeroides: Tran­scriptional regulation of the heat-induced groESL operon. J. BacterioL 179, 487-495.
• 23. Wetzstein, M., Volker, U., Dedio, J., Lobau, S., Zuber, U., Schiesswohl, M., Herget, C., Hec- kcr, M. & Schumann, W. (1992) Cloning, se­quencing, and molecular analysis of the dnaK locus from Bacillus subtilis. J. BacterioL 174, 3300-3310.
• 24. Narberhaus, F., Giebeler, K. & Bahl, H. (1992) Molecular characterization of the dnaK gene region of Clostridium acetobutylicum, including grpE, dnaJ, and a new heat shock gene. J. Bac­terioL 174, 3290-3299.
• 25. Cellier, M.F.M., Teyssier, J., Nicolas, M., Li- autard, J.P., Marti, J. & Sri Widada, J. (1992) Cloning and characterization of the Brucella ovis heat shock protein DnaK functionally ex­pressed in Escherichia coli. J. BacterioL 174, 8036-8042.
• 26. Tilly, K., Hauser, R., Campbell, J. & Osth- eimer, G.J. (1993) Isolation of dnaJ, dnaK, and GrpE homologues from Borrelia burgdor­feri and complementation of Escherichia coli mutants. Mol. Microbiol. 7, 359-369.
• 27. Minder, A.C., Narberhaus, F., Babst, M., Hen- necke, H. & Fischer, H.-M. (1997) The dnaKJ operon belongs to the a32 -dependent class of heat shock genes in Bradyrhizobium japoni- cum. Mol Gen. Genet. 254, 195-206.
• 28. Chitnis, P.R. & Nelson, N. (1991) Molecular cloning of the genes encoding two chaperone proteins of the cyanobacterium Synechocystis sp. PCC 80S. J. Biol Chem. 266, 58-65.
• 29. Webb, R., Reddy, K.J. & Sherman, L.A. (1990) Regulation and sequence of the Synechococcus sp. strain PCC 7942 groESL operon, encoding a cyanobacterial chaperonin. J. Bacteriol. 172, 5079-5088.
• 30. Michel, G.P. (1993) Cloning and expression in Escherichia coli of the dnaK gene of Zymonas mobilis. J. Bacteriol 175, 3228-3231.
• 31. Zuber, M., Hoover, T.A., Dertzbaugh. M.T. & Court, D.L. (1995) Analysis of the DnaK mo­lecular chaperone system of Francisella tular­ensis. Gene 164, 149-152.
• 32. Miyazaki, T., Tanaka, S., Fujita, H. & Itikawa, H. (1992) DNA sequence analysis of the dnaK gene of Escherichia coli B and two dnaK genes carrying the temperature-sensitive mutations dnaK7 (Ts) and dnaK756£T&). J. Bacteriol 174, 3715-3722.
• 33.1vic, A., Olden, D., Wallington, E.J. & Lund, P.A. (1997) Deletion of Escherichia coli groEL is complemented by Rhizobium leguminosarum groEL homologue at 37°C but not at 43°C. Gene 194, 1-8.
• 34. Frydman, J. & Hartl. F.U. (1994) Molecular chaperone functions of hsp70 and hsp60 in protein folding; in The Biology of Heat Shock Proteins and Molecular Chaperones (Morimoto, R.I., Tissieres, A. & Georgopoulos, C., eds.) pp. 251-283, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
• 35. Fri8chauf, A.-M., Lehrach, H., Poustka, A. & Murray, N. (1983) Lambda replacement vec­tors carrying poly linker sequences. J. Mol Biol. 170, 827-842.
• 36. Fayet, 0., Louam, J.M. & Georgopoulos, C. (1986) Suppression of Escherichia coli dnaA46 mutation by amplification of groES and groEL genes. Mol Gen. Genet. 202, 435-445.
• 37. Hemmingsen, S.M., Woolford, C., Tilly, K., van der Vies, S.M. & Georgopoulos, C. (1988) Homologous plant and bacterial proteins chap­erone oligomeric protein assembly. Nature 333, 330-334.