Molecular cloning and nucleotide sequence of a cDNA encoding recoverin from human retina.
In the process of photoreceptor signal transduction, light initiates an enzymatic cascade that leads to hydrolysis of cyclic GMP (cGMP) and closure of cGMP-gated sodium-calcium channels resulting in photoreceptor hyperpolarization. Recoverin is a calcium-binding protein that is thought to reverse the effects of light on cGMP levels by activating guanylate cyclase. Guanylate cyclase produces cGMP to overcome the cGMP-hydrolysing effect of phosphodiesterase, and reopens the sodium-calcium channels in photoreceptor outer segments. We have cloned and sequenced a cDNA encoding recoverin in human retina. The human nucleotide sequence is 88% identical to the bovine sequence, and contains a 600-base pair (bp) open reading frame encoding 200 amino acids. In situ hybridization of cultured Y79 human retinoblastoma cells with a radioactive recoverin cDNA probe showed intense, specific labeling of the cytoplasm, indicating the presence of mRNA encoding recoverin. Direct sequencing of a Y79 retinoblastoma cDNA polymerase chain reaction (PCR) product confirmed the presence of recoverin in this human cell line.
Department of Neurobiology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, NC.
A F
AF
Wiechmann
J A
JA
Hammarback
D000595:Amino Acid Sequence; D000951:Antigens, Neoplasm / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D005136:Eye Proteins / Q000235:genetics*; D051597:Hippocalcin; D006801:Humans; D017403:In Situ Hybridization; D008074:Lipoproteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D051603:Recoverin; D012160:Retina; D012175:Retinoblastoma / Q000235:genetics
D000951:Antigens, Neoplasm; D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; C496731:RCVRN protein, human; D012333:RNA, Messenger; D051603:Recoverin; D051597:Hippocalcin