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Volume 272, Number 17,
Issue of April 25, 1997
pp. 11479-11486
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
gRICH68 and gRICH70 Are 2 ,3 -Cyclic-nucleotide
3 -Phosphodiesterases Induced during Goldfish Optic Nerve
Regeneration
(Received for publication, December 31, 1996, and in revised form, February 12, 1997)
Rafael P.
Ballestero
,
George R.
Wilmot
,
Bernard W.
Agranoff
and
Michael D.
Uhler
From the Department of Biological Chemistry and Mental Health
Research Institute, University of Michigan,
Ann Arbor, Michigan 48109
Biochemical characterization of changes in gene
expression that accompany optic nerve regeneration has led to the
identification of proteins that may play key roles in the regeneration
process. In this report, a cDNA encoding gRICH70, a novel isoform
of the regeneration-induced gRICH68 protein, has been identified and characterized in goldfish. Both gRICH68 and gRICH70 show significant homology (34-36%) to mammalian 2 ,3 -cyclic-nucleotide
3 -phosphodiesterases (CNPases), hence the name oldfish
egeneration-nduced NPase omolog (gRICH). The predicted 431-amino acid gRICH70
protein is 88% homologous to gRICH68, and the retinal mRNA for
gRICH70 is coordinately induced with gRICH68 mRNA during optic
nerve regeneration. Enzymatic analysis of recombinant proteins confirms
that both gRICH proteins possess CNPase activity. Despite the
relatively limited sequence homology, the kinetic constants obtained
suggest that both gRICH proteins are at least as efficient as
recombinant mouse CNP1 in catalyzing the hydrolysis of 2 ,3 -cAMP.
Immunoprecipitation studies indicate that gRICH proteins are
responsible for the majority of the CNPase activity detected in
regenerating goldfish retinas. The evidence presented demonstrates that
gRICH68 and gRICH70 correspond to a previously described doublet of
acidic proteins that are selectively induced in the goldfish retina
during optic nerve regeneration. Thus, CNPase enzyme activity is
implicated for the first time in the process of nerve regeneration.

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Copyright © 1997 by the American Society for Biochemistry and Molecular Biology.
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