title: Sensory rhodopsins I and II modulate a methylation/demethylation system in Halobacterium halobium phototaxis.
abstract: This work demonstrates that phototaxis stimuli in the archaebacterium Halobacterium halobium control a methylation/demethylation system in vivo through photoactivation of sensory rhodopsin I (SR-I) in either its attractant or repellent signaling form as well as through the repellent receptor sensory rhodopsin II (SR-II, also called phoborhodopsin). The effects of positive stimuli that suppress swimming reversals (i.e., an increase in attractant or decrease in repellent light) and negative stimuli that induce swimming reversals (i.e., a decrease in attractant or increase in repellent light) through each photoreceptor were monitored by assaying release of volatile [3H]methyl groups. This assay has been used to measure [3H]methanol produced during the process of adaptation to chemotactic stimuli in eubacteria. In H. halobium positive photostimuli produce a transient increase in the rate of demethylation followed by a decrease below the unstimulated value, whereas negative photostimuli cause an increase followed by a rate similar to that of the unstimulated value. Photoactivation of the SR-I attractant and simultaneous photoactivation of the SR-II repellent receptors cancel in their effects on demethylation, demonstrating the methylation system is regulated by an integrated signal. Analysis of mutants indicates that the source for the volatile methyl groups is intrinsic membrane proteins distinct from the chromoproteins that share the membrane. A methyl-accepting protein (94 kDa) previously correlated in amount with the SR-I chromoprotein (25 kDa) is shown here to be missing in a recently isolated SR-I-SR-II+ mutant (Flx3b), thus confirming the association of this protein with SR-I. Photoactivated SR-II in mutant Flx3b controls demethylation, predicting the existence of a photomodulated methyl-accepting component distinct from the 94-kDa protein of SR-I. We present a model in which the three known phototaxis signaling receptor states (the attractant receptor SR-I587, its repellent form S373, and the repellent receptor SR-II490) are coupled to two distinct transducers the demethylation of which is controlled by one integrated signal.
authors.affiliation: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461.
authors.forename: E N
authors.initials: EN
authors.lastname: Spudich
authors.forename: T
authors.initials: T
authors.lastname: Takahashi
authors.forename: J L
authors.initials: JL
authors.lastname: Spudich
mesh_terms: D019843:Archaeal Proteins; D001436:Bacteriorhodopsins / Q000378:metabolism*; D002338:Carotenoids; D002465:Cell Movement; D006217:Halobacterium / Q000502:physiology*; D025602:Halorhodopsins; D007700:Kinetics; D008027:Light; D008715:Methionine / Q000378:metabolism*; D008745:Methylation; D008962:Models, Theoretical; D011865:Radioisotope Dilution Technique; D012172:Retinaldehyde / Q000378:metabolism; D025603:Sensory Rhodopsins; D013997:Time Factors; D014316:Tritium
chemical_list: D019843:Archaeal Proteins; D025602:Halorhodopsins; C061497:SRI protein, Halobacterium; D025603:Sensory Rhodopsins; C061508:sensory rhodopsin II protein, archaeal; D014316:Tritium; D002338:Carotenoids; D001436:Bacteriorhodopsins; D008715:Methionine; D012172:Retinaldehyde