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. 1994 May 1;13(9):2150–2155. doi: 10.1002/j.1460-2075.1994.tb06491.x

Phototaxis of Halobacterium salinarium requires a signalling complex of sensory rhodopsin I and its methyl-accepting transducer HtrI.

M Krah 1, W Marwan 1, A Verméglio 1, D Oesterhelt 1
PMCID: PMC395068  PMID: 8187768

Abstract

Sensory rhodopsin I (SRI) is a photoreceptor that mediates phototaxis in the archaeon Halobacterium salinarium. Receptor excitation is relayed to the motility system of the cell by the methyl-accepting transducer protein HtrI. In membranes prepared from cells that lack HtrI the absorbance difference maximum of SRI was shifted from 587 to 565 nm. The thermal decay of the metastable photocycle intermediate SRI373 was measured as time-dependent recovery of the absorbance at 590 nm. In the absence of HtrI the decay was slowed down by two orders of magnitude. When SRI was overproduced in cells that contained normal levels of HtrI, the decay of SRI373 was biexponential indicating two kinetically distinct species. Spectroscopic measurements on intact cells revealed the same effect of HtrI on SRI photocycling as found in isolated membranes. By transient exposure of membranes from wild-type cells to low ionic strength, the decay of SR373 was slowed to the same value found for untreated membranes in the absence of HtrI. In parallel, the absorbance difference maximum was shifted to 565 nm indicating that a physical interaction of HtrI and SRI had been irreversibly destroyed. Overproduction of SRI in the presence of wild-type amounts of HtrI did not increase the light sensitivity of the cells to orange light step down stimulation. It is concluded that SRI and HtrI form a stable complex in the cell membrane that signals to the flagellar motor and defines absorbance maximum, photocycling rate and photochemical efficiency of SRI.

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Selected References

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  1. Alam M., Hazelbauer G. L. Structural features of methyl-accepting taxis proteins conserved between archaebacteria and eubacteria revealed by antigenic cross-reaction. J Bacteriol. 1991 Sep;173(18):5837–5842. doi: 10.1128/jb.173.18.5837-5842.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alam M., Lebert M., Oesterhelt D., Hazelbauer G. L. Methyl-accepting taxis proteins in Halobacterium halobium. EMBO J. 1989 Feb;8(2):631–639. doi: 10.1002/j.1460-2075.1989.tb03418.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alam M., Oesterhelt D. Morphology, function and isolation of halobacterial flagella. J Mol Biol. 1984 Jul 15;176(4):459–475. doi: 10.1016/0022-2836(84)90172-4. [DOI] [PubMed] [Google Scholar]
  4. Blanck A., Oesterhelt D., Ferrando E., Schegk E. S., Lottspeich F. Primary structure of sensory rhodopsin I, a prokaryotic photoreceptor. EMBO J. 1989 Dec 20;8(13):3963–3971. doi: 10.1002/j.1460-2075.1989.tb08579.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bogomolni R. A., Spudich J. L. Identification of a third rhodopsin-like pigment in phototactic Halobacterium halobium. Proc Natl Acad Sci U S A. 1982 Oct;79(20):6250–6254. doi: 10.1073/pnas.79.20.6250. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ehrlich B. E., Schen C. R., Spudich J. L. Bacterial rhodopsins monitored with fluorescent dyes in vesicles and in vivo. J Membr Biol. 1984;82(1):89–94. doi: 10.1007/BF01870735. [DOI] [PubMed] [Google Scholar]
  7. Ferrando-May E., Brustmann B., Oesterhelt D. A C-terminal truncation results in high-level expression of the functional photoreceptor sensory rhodopsin I in the archaeon Halobacterium salinarium. Mol Microbiol. 1993 Sep;9(5):943–953. doi: 10.1111/j.1365-2958.1993.tb01224.x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Hildebrand E., Dencher N. Two photosystems controlling behavioural responses of Halobacterium halobium. Nature. 1975 Sep 4;257(5521):46–48. doi: 10.1038/257046a0. [DOI] [PubMed] [Google Scholar]
  10. Manor D., Hasselbacher C. A., Spudich J. L. Membrane potential modulates photocycling rates of bacterial rhodopsins. Biochemistry. 1988 Aug 9;27(16):5843–5848. doi: 10.1021/bi00416a004. [DOI] [PubMed] [Google Scholar]
  11. Marwan W., Oesterhelt D. Quantitation of photochromism of sensory rhodopsin-I by computerized tracking of Halobacterium halobium cells. J Mol Biol. 1990 Sep 20;215(2):277–285. doi: 10.1016/S0022-2836(05)80346-8. [DOI] [PubMed] [Google Scholar]
  12. Marwan W., Schäfer W., Oesterhelt D. Signal transduction in Halobacterium depends on fumarate. EMBO J. 1990 Feb;9(2):355–362. doi: 10.1002/j.1460-2075.1990.tb08118.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Montrone M., Marwan W., Grünberg H., Musseleck S., Starostzik C., Oesterhelt D. Sensory rhodopsin-controlled release of the switch factor fumarate in Halobacterium salinarium. Mol Microbiol. 1993 Dec;10(5):1077–1085. doi: 10.1111/j.1365-2958.1993.tb00978.x. [DOI] [PubMed] [Google Scholar]
  14. Oesterhelt D., Krippahl G. Phototrophic growth of halobacteria and its use for isolation of photosynthetically-deficient mutants. Ann Microbiol (Paris) 1983 Jul-Aug;134B(1):137–150. doi: 10.1016/s0769-2609(83)80101-x. [DOI] [PubMed] [Google Scholar]
  15. Oesterhelt D., Stoeckenius W. Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol. 1974;31:667–678. doi: 10.1016/0076-6879(74)31072-5. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Schegk E. S., Oesterhelt D. Isolation of a prokaryotic photoreceptor: sensory rhodopsin from halobacteria. EMBO J. 1988 Sep;7(9):2925–2933. doi: 10.1002/j.1460-2075.1988.tb03151.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schimz A. Methylation of membrane proteins is involved in chemosensory and photosensory behavior of Halobacterium halobium. FEBS Lett. 1981 Mar 23;125(2):205–207. doi: 10.1016/0014-5793(81)80719-3. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. 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]
  22. 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]
  23. Spudich J. L., Bogomolni R. A. Sensory rhodopsin I: receptor activation and signal relay. J Bioenerg Biomembr. 1992 Apr;24(2):193–200. doi: 10.1007/BF00762677. [DOI] [PubMed] [Google Scholar]
  24. Sundberg S. A., Alam M., Lebert M., Spudich J. L., Oesterhelt D., Hazelbauer G. L. Characterization of Halobacterium halobium mutants defective in taxis. J Bacteriol. 1990 May;172(5):2328–2335. doi: 10.1128/jb.172.5.2328-2335.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Uhl R., Meyer B., Desel H. A polychromatic flash photolysis apparatus (PFPA). J Biochem Biophys Methods. 1984 Nov;10(1-2):35–48. doi: 10.1016/0165-022x(84)90048-4. [DOI] [PubMed] [Google Scholar]
  27. Yan B., Takahashi T., Johnson R., Derguini F., Nakanishi K., Spudich J. L. All-trans/13-cis isomerization of retinal is required for phototaxis signaling by sensory rhodopsins in Halobacterium halobium. Biophys J. 1990 Apr;57(4):807–814. doi: 10.1016/S0006-3495(90)82600-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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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