Isolation of cDNA clones encoding RICH: a protein induced during goldfish optic nerve regeneration with homology to mammalian 2',3'-cyclic-nucleotide 3'-phosphodiesterases.
Using data derived from peptide sequencing of p68/70, a protein doublet induced during optic nerve regeneration in goldfish, we have isolated cDNAs that encode RICH (regeneration-induced CNPase homolog) from a goldfish regenerating retina cDNA library. The predicted RICH protein comprises 411 amino acids, possesses a pI of 4.48, and shows significant homology to the mammalian myelin marker enzyme 2',3'-cyclic-nucleotide 3'-phosphodiesterase (CNPase; EC 3.1.4.37). The mRNA encoding RICH was demonstrated, by both Northern blot analysis and RNase protection assays, to be induced as much as 8-fold in regenerating goldfish retinas at 20 days after nerve crush. Analysis of total RNA samples from various tissues showed a broad distribution of RICH mRNA, with the highest levels observed in gravid ovary. The data obtained strongly suggest that RICH is identical or very similar to p68/70. The molecular cloning of RICH provides the means for a more detailed analysis of its function in nerve regeneration. Additionally, the homology of RICH and CNPase suggests that further investigation may provide additional insight into the role of these proteins in the nervous system.
Ballestero
R P
RP
Department of Biological Chemistry, University of Michigan, Ann Arbor 48109, USA.
Wilmot
G R
GR
Leski
M L
ML
Uhler
M D
MD
Agranoff
B W
BW
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004790:Enzyme Induction; D029941:Fish Proteins; D018507:Gene Expression Regulation, Developmental; D006054:Goldfish; D008322:Mammals / Q000235:genetics; D008969:Molecular Sequence Data; D009416:Nerve Regeneration / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009900:Optic Nerve / Q000201:enzymology / Q000502:physiology*; D010727:Phosphoric Diester Hydrolases; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution
D018076:DNA, Complementary; D029941:Fish Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; C095683:G-rich protein, Carassius auratus; D010727:Phosphoric Diester Hydrolases