In vivo characterization of phytochrome-phycocyanobilin adducts in yeast.
The in vivo reconstitution of phycocyanobilin with apophytochrome leads to photoreversible adducts in living yeast cells. Investigations with the rice phytochrome A phycocyanobilin adduct (PHYA*) and the tobacco phytochrome B phycocyanobilin adduct (PHYB*) show that the protein stability in yeast is independent of the form of the photoreceptor. After in vivo assembly and irradiation with red light, 25.6% of the far-red light-absorbing form of PHYB* exhibited dark reversion with a half-life time of approximately 20 min. Control experiments with PHYA* revealed no dark reversion. The data indicate that the molecular basis for this reaction is the formation of heterodimers between the red and the far-red light absorbing form of phytochrome. Electron microscopic in situ localizations and in vitro sequestering experiments showed that phytochrome A was able to sequester in yeast. On the electron microscopic level, the sequestered areas of phytochrome from etiolated plants and yeast are indistinguishable. The sequestering reaction in yeast is independent of the formation of the far-red light absorbing form of phytochrome. Therefore, we discuss a new model for this reaction in plants. Since it is unlikely that yeast cells contain elements that distinguish between phytochrome A and B, we conclude that sequestering and dark reversion reflect intrinsic properties of phytochrome.
Kunkel
T
T
Institut für Biologie II, Albert-Ludwigs-Universität Freiburg, Federal Republic of Germany.
Speth
V
V
Büche
C
C
Schäfer
E
E
D015966:Gene Expression Regulation, Fungal; D018506:Gene Expression Regulation, Plant; D008027:Light; D016253:Microscopy, Immunoelectron; D010786:Photoreceptor Cells / Q000378:metabolism / Q000528:radiation effects; D052980:Phycobilins; D010798:Phycocyanin / Q000032:analysis / Q000378:metabolism* / Q000528:radiation effects; D010834:Phytochrome / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D010944:Plants / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D011758:Pyrroles / Q000032:analysis / Q000378:metabolism* / Q000528:radiation effects; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D045725:Tetrapyrroles
D052980:Phycobilins; D011758:Pyrroles; D045725:Tetrapyrroles; D010798:Phycocyanin; D010834:Phytochrome; C011069:phycocyanobilin