Structure of the 55-kDa regulatory subunit of protein phosphatase 2A: evidence for a neuronal-specific isoform.
The trimeric form of protein phosphatase 2A (PP2A1 or polycation-stimulated protein phosphatase H1) was purified to homogeneity from rabbit skeletal muscle. Preparative SDS-polyacrylamide gel electrophoresis was used to purify the individual subunits with relative molecular masses of 36, 55, and 65 kDa. Sequence analysis of five peptides from the 65-kDa regulatory subunit (PR65) suggested that it was identical with the PR65 subunit derived from the dimeric protein phosphatase 2A2. Amino acid sequences derived from the 55-kDa regulatory subunit (PR55) were used to clone human and rabbit cDNAs encoding this protein. The PR55 subunit was found to be encoded by two genes, termed alpha and beta. The open reading frames of the PR55 alpha and beta cDNAs spanned 1341 and 1329 nucleotides, respectively, and predicted proteins with a molecular mass of about 52 kDa that are 86% identical. Comparison of the human PR55 amino acid sequences with the data obtained from the rabbit skeletal muscle protein and a partial rabbit PR55 beta cDNA clone indicated a high degree of conservation. Analysis of the mRNA expression in human cell lines revealed that the PR55 alpha isoform was encoded by two transcripts of about 2.3 and 2.5 kb and a less abundant 4.4-kb mRNA. Whereas a PR55 beta transcript of about 2.3 kb was detected at high levels in the neuroblastoma derived cell line LA-N-1, the level of the mRNA was very low in the other human cell lines analyzed. Interestingly, the PR55 sequence showed limited homology to the catalytic domain (domains VI-IX) of the c-abl protein tyrosine kinase.
Mayer
R E
RE
Friedrich Miescher-Institut, Basel, Switzerland.
Hendrix
P
P
Cron
P
P
Matthies
R
R
Stone
S R
SR
Goris
J
J
Merlevede
W
W
Hofsteenge
J
J
Hemmings
B A
BA
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D015870:Gene Expression; D015698:Genomic Library; D006801:Humans; D007536:Isomerism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009474:Neurons / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000737:chemistry / Q000235:genetics*; D054648:Protein Phosphatase 2; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D016315:Proto-Oncogene Proteins c-abl / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D012333:RNA, Messenger; D004247:DNA; D011505:Protein-Tyrosine Kinases; D016315:Proto-Oncogene Proteins c-abl; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2