Retinoylation of the cAMP-binding regulatory subunits of type I and type II cAMP-dependent protein kinases in HL60 cells.
Retinoylation (retinoic acid acylation) is a post-translational modification of proteins occurring in a variety of eukaryotic cell lines. There are at least 20 retinoylated proteins in the human myeloid leukemia cell line HL60 (N. Takahashi and T.R. Breitman (1990) J. Biol. Chem. 265, 19, 158-19, 162). Here we found that some retinoylated proteins may be cAMP-binding proteins. Five proteins, covalently labeled by 8-azido-[32P]cAMP which specifically reacts with the regulatory subunits of cAMP-dependent protein kinase, comigrated on two-dimensional polyacrylamide gel electrophoresis with retinoylated proteins of Mr 37,000 (p37RA), 47,000 (p47RA), and 51,000 (p51RA) labeled by [3H]retinoic acid treatment of intact cells. Furthermore, p47RA coeluted on Mono Q anion exchange chromatography with the type I cAMP-dependent protein kinase holoenzyme and p51RA coeluted on Mono Q anion exchange chromatography with the type II cAMP-dependent protein kinase holoenzyme. An antiserum specific to RI, the cAMP-binding regulatory subunit of type I cAMP-dependent protein kinase, immunoprecipitated p47RA. An antiserum specific to RII, the cAMP-binding regulatory subunit of type II cAMP-dependent protein kinase, immunoprecipitated p51RA. These results indicate that both the RI and the RII regulatory subunits of cAMP-dependent protein kinase are retinoylated. Thus, an early event in RA-induced differentiation of HL60 cells may be the retinoylation of subpopulations of both RI and RII.
Takahashi
N
N
Laboratory of Biological Chemistry, National Cancer Institute, Bethesda, Maryland 20892.
Liapi
C
C
Anderson
W B
WB
Breitman
T R
TR
D001386:Azides / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry*; D002460:Cell Line; D002852:Chromatography, Ion Exchange; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000737:chemistry* / Q000378:metabolism; D002373:Cyclic AMP Receptor Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007951:Leukemia, Myeloid / Q000201:enzymology*; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000737:chemistry*; D011233:Precipitin Tests; D011494:Protein Kinases / Q000737:chemistry*; D012965:Sodium Chloride; D014212:Tretinoin / Q000737:chemistry*
D001386:Azides; D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D009363:Neoplasm Proteins; C017042:8-azidoadenosine-3',5'-monophosphate; D012965:Sodium Chloride; D014212:Tretinoin; D000242:Cyclic AMP; D011494:Protein Kinases