The abundant retinal protein of the Chlamydomonas eye is not the photoreceptor for phototaxis and photophobic responses.
The chlamyopsin gene (cop) encodes the most abundant eyespot protein in the unicellular green alga Chlamydomonas reinhardtii. This opsin-related protein (COP) binds retinal and was thought to be the photoreceptor controlling photomovement responses via a set of photoreceptor currents. Unfortunately, opsin-deficient mutants are not available and targeted disruption of non-selectable nuclear genes is not yet possible in any green alga. Here we show that intron-containing gene fragments directly linked to their intron-less antisense counterpart provide efficient post-transcriptional gene silencing (PTGS) in C. reinhardtii, thus allowing an efficient reduction of a specific gene product in a green alga. In opsin-deprived transformants, flash-induced photoreceptor currents (PC) are left unchanged. Moreover, photophobic responses as studied by motion analysis and phototaxis tested in a light-scattering assay were indistinguishable from the responses of untransformed wild-type cells. We conclude that phototaxis and photophobic responses in C. reinhardtii are triggered by an as yet unidentified rhodopsin species.
Fuhrmann
M
M
Institut für Biochemie I, Universität Regensburg, 93040 Regensburg, Germany. markus.fuhrmann@vkl.uni-regensburg.de
Stahlberg
A
A
Govorunova
E
E
Rank
S
S
Hegemann
P
P
D020418:Algal Proteins; D000818:Animals; D001059:Apoproteins / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D020868:Gene Silencing; D047908:Intracellular Signaling Peptides and Proteins; D008027:Light; D010786:Photoreceptor Cells / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000032:analysis; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D012160:Retina / Q000378:metabolism*; D017299:Rod Opsins / Q000378:metabolism*; D014170:Transformation, Genetic
D020418:Algal Proteins; D001059:Apoproteins; D002352:Carrier Proteins; D005136:Eye Proteins; D047908:Intracellular Signaling Peptides and Proteins; D015800:Protozoan Proteins; D016337:RNA, Catalytic; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D017299:Rod Opsins; C097338:chlamyopsin protein, Chlamydomonas reinhardtii; C418481:hairpin ribozyme; D012313:RNA