ARPP-21, a cyclic AMP-regulated phosphoprotein enriched in dopamine-innervated brain regions. II. Molecular cloning and nucleotide sequence.
A cDNA clone for the mRNA of bovine ARPP-21 (cAMP-regulated phosphoprotein, Mr = 21,000 as determined by SDS-PAGE) was isolated from a modified Okayama-Berg plasmid library. Transformed Escherichia coli colonies were screened by in situ colony hybridization with 2 different oligonucleotide probes derived from the amino acid sequence of the bovine protein. Sequence analysis of the longest cDNA clone, pTKAI [2407 nucleotides plus a poly(A) tail], revealed a 267-nucleotide-long coding region in agreement with the bovine ARPP-21 amino acid sequence (Williams et al., 1989). Southern blot analysis of total bovine genomic DNA raised the possibility that there may be 2 genes coding for ARPP-21. Northern blot analysis of total cellular RNA from bovine caudate nucleus and other brain regions demonstrated the existence of 2 major mRNA species, 2.5 and 1.0 kb in length, probably derived from use of alternate polyadenylation sites. There was a differential expression of these 2 mRNAs within the brain. Both ARPP-21 mRNAs were most abundant in the caudate nucleus, where the concentration of the protein is highly enriched.
Kurihara
T
T
Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021.
Ehrlich
M E
ME
Horiuchi
J
J
Nasu
T
T
Greengard
P
P
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000502:physiology*; D004247:DNA / Q000235:genetics; D004298:Dopamine / Q000502:physiology*; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics
D010750:Phosphoproteins; D012333:RNA, Messenger; C061368:cyclic AMP-regulated phosphoprotein ARPP-21; D004247:DNA; D000242:Cyclic AMP; D004298:Dopamine