Structure and Ca2+ regulation of frog photoreceptor guanylate cyclase, ROS-GC1.
Rod outer segment membrane guanylate cyclase (ROS-GC) is a critical component of the vertebrate phototransduction machinery. In response to photoillumination, it senses a decline in free Ca(2+) levels from 500 to below 100 nM, becomes activated, and replenishes the depleted cyclic GMP pool to restore the dark state of the photoreceptor cell. It exists in two forms, ROS-GC1 and ROS-GC2. In outer segments, ROS-GCs sense fluctuations in Ca(2+) via two Ca(2+)-binding proteins, which have been termed GCAP1 and GCAP2. In the present study we report on the cloning of two ROS-GCs from the frog retinal cDNA library. These cyclases are the structural and functional counterparts of the mammalian ROS-GC1 and ROS-GC2. There is, however, an important difference between the regulation of mammalian and frog ROS-GC1: In contrast to the mammalian, the frog form does not require the myristoylated form of GCAP1 for its Ca(2+)-dependent modulation. This feature is not dependent upon the ability of frog GCAP1 to bind Ca(2+) because unmyristoylated GCAP1 mutants which do not bind Ca(2+), activate frog ROS-GC1. The findings establish frog as a suitable phototransduction model and show a facet of frog ROS-GC signaling, which is not shared by the mammalian form.
Subbaraya
Iswari
I
Department of Ophthalmology, Baylor College of Medicine, Houston, TX, USA.
Zhao
Chong
C
Duda
Teresa
T
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D006152:Cyclic GMP / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D015723:Gene Library; D006162:Guanylate Cyclase / Q000096:biosynthesis* / Q000378:metabolism / Q000502:physiology*; D007313:Insecta; D008969:Molecular Sequence Data; D009154:Mutation; D009227:Myristic Acids / Q000378:metabolism; D011485:Protein Binding; D011898:Ranidae; D011956:Receptors, Cell Surface; D012374:Rod Cell Outer Segment / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D014162:Transfection; D014785:Vision, Ocular
D018076:DNA, Complementary; D009227:Myristic Acids; D011956:Receptors, Cell Surface; C110715:guanylate cyclase 1; D006162:Guanylate Cyclase; D006152:Cyclic GMP; D002118:Calcium