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. 1994 Nov;176(22):6931–6935. doi: 10.1128/jb.176.22.6931-6935.1994

Identification of distinct domains for signaling and receptor interaction of the sensory rhodopsin I transducer, HtrI.

V J Yao 1, E N Spudich 1, J L Spudich 1
PMCID: PMC197063  PMID: 7961454

Abstract

The phototaxis-deficient mutant of Halobacterium salinarium, Pho81, lacks both sensory rhodopsin I (SR-I) and its putative transducer protein HtrI, according to immunoblotting and spectroscopic criteria. From restriction analysis and selected DNA sequencing, we have determined that the SR-I- HtrI- phenotype results from an insertion of a 520-bp transposable element, ISH2, into the coding region of the SR-I apoprotein gene sopI and deletion of 11 kbp upstream of ISH2 including the first 164 bp of sopI and the entire htrI gene. SR-I and HtrI expression as well as full phototaxis sensitivity are restored by transformation with a halobacterial plasmid carrying the htrI-sopI gene pair and their upstream promoter region. An internal deletion of a portion of htrI encoding the putative methylation and signaling domains of HtrI (253 residues) prevents the restoration of phototaxis, providing further evidence for the role of HtrI as a transducer for SR-I. Analysis of flash-induced photochemical reactions of SR-I over a range of pH shows that the partially deleted HtrI maintains SR-I interactions sites responsible for modulation of the SR-I photocycle.

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Selected References

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  1. Borkovich K. A., Simon M. I. Coupling of receptor function to phosphate-transfer reactions in bacterial chemotaxis. Methods Enzymol. 1991;200:205–214. doi: 10.1016/0076-6879(91)00140-r. [DOI] [PubMed] [Google Scholar]
  2. Cline S. W., Doolittle W. F. Efficient transfection of the archaebacterium Halobacterium halobium. J Bacteriol. 1987 Mar;169(3):1341–1344. doi: 10.1128/jb.169.3.1341-1344.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. DasSarma S., RajBhandary U. L., Khorana H. G. High-frequency spontaneous mutation in the bacterio-opsin gene in Halobacterium halobium is mediated by transposable elements. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2201–2205. doi: 10.1073/pnas.80.8.2201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunn R., McCoy J., Simsek M., Majumdar A., Chang S. H., Rajbhandary U. L., Khorana H. G. The bacteriorhodopsin gene. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6744–6748. doi: 10.1073/pnas.78.11.6744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ferrando-May E., Krah M., Marwan W., Oesterhelt D. The methyl-accepting transducer protein HtrI is functionally associated with the photoreceptor sensory rhodopsin I in the archaeon Halobacterium salinarium. EMBO J. 1993 Aug;12(8):2999–3005. doi: 10.1002/j.1460-2075.1993.tb05968.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Krah M., Marwan W., Verméglio A., Oesterhelt D. Phototaxis of Halobacterium salinarium requires a signalling complex of sensory rhodopsin I and its methyl-accepting transducer HtrI. EMBO J. 1994 May 1;13(9):2150–2155. doi: 10.1002/j.1460-2075.1994.tb06491.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Krebs M. P., Hauss T., Heyn M. P., RajBhandary U. L., Khorana H. G. Expression of the bacterioopsin gene in Halobacterium halobium using a multicopy plasmid. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):859–863. doi: 10.1073/pnas.88.3.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Krebs M. P., Mollaaghababa R., Khorana H. G. Gene replacement in Halobacterium halobium and expression of bacteriorhodopsin mutants. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1987–1991. doi: 10.1073/pnas.90.5.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Krebs M. P., Spudich E. N., Khorana H. G., Spudich J. L. Synthesis of a gene for sensory rhodopsin I and its functional expression in Halobacterium halobium. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3486–3490. doi: 10.1073/pnas.90.8.3486. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lam W. L., Doolittle W. F. Shuttle vectors for the archaebacterium Halobacterium volcanii. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5478–5482. doi: 10.1073/pnas.86.14.5478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olson K. D., Spudich J. L. Removal of the transducer protein from sensory rhodopsin I exposes sites of proton release and uptake during the receptor photocycle. Biophys J. 1993 Dec;65(6):2578–2585. doi: 10.1016/S0006-3495(93)81295-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  14. Spudich E. N., Hasselbacher C. A., Spudich J. L. Methyl-accepting protein associated with bacterial sensory rhodopsin I. J Bacteriol. 1988 Sep;170(9):4280–4285. doi: 10.1128/jb.170.9.4280-4285.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Spudich E. N., Spudich J. L. The photochemical reactions of sensory rhodopsin I are altered by its transducer. J Biol Chem. 1993 Aug 5;268(22):16095–16097. [PubMed] [Google Scholar]
  16. Spudich E. N., Sundberg S. A., Manor D., Spudich J. L. Properties of a second sensory receptor protein in Halobacterium halobium phototaxis. Proteins. 1986 Nov;1(3):239–246. doi: 10.1002/prot.340010306. [DOI] [PubMed] [Google Scholar]
  17. Spudich E. N., Takahashi T., Spudich J. L. Sensory rhodopsins I and II modulate a methylation/demethylation system in Halobacterium halobium phototaxis. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7746–7750. doi: 10.1073/pnas.86.20.7746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Spudich J. L., Bogomolni R. A. Mechanism of colour discrimination by a bacterial sensory rhodopsin. Nature. 1984 Dec 6;312(5994):509–513. doi: 10.1038/312509a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sundberg S. A., Alam M., Spudich J. L. Excitation signal processing times in Halobacterium halobium phototaxis. Biophys J. 1986 Nov;50(5):895–900. doi: 10.1016/S0006-3495(86)83530-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sundberg S. A., Bogomolni R. A., Spudich J. L. Selection and properties of phototaxis-deficient mutants of Halobacterium halobium. J Bacteriol. 1985 Oct;164(1):282–287. doi: 10.1128/jb.164.1.282-287.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  22. Yao V. J., Spudich J. L. Primary structure of an archaebacterial transducer, a methyl-accepting protein associated with sensory rhodopsin I. Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):11915–11919. doi: 10.1073/pnas.89.24.11915. [DOI] [PMC free article] [PubMed] [Google Scholar]

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