Selective regulation of expression of protein kinase C beta isoenzymes occurs via alternative splicing.
The mechanisms involved in regulating the selective expression of protein kinase C (PKC) isoenzymes are poorly understood. Two human B lymphoblastoid cell lines, IM-9 and BJA-B, exhibited differential expression of the two alternatively spliced products of the PKC beta gene, PKC beta I and beta II. The IM-9 cell line expressed 3-4-fold more PKC beta II protein than the BJA-B cell line, whereas the BJA-B cell line expressed 2-3-fold more PKC beta I protein. This differential expression was found to be regulated at the mRNA level. Comparison of PKC beta I and beta II messages in poly(A)+ mRNA and total cellular RNA revealed that selective polyadenylation was not involved. The messages for PKC beta I and beta II had comparable half-lives in both cell lines, ruling out differential message stability. In addition, similar ratios of PKC beta I and beta II messages in cytosolic and nuclear fractions suggested that differential mRNA transport was not involved. In the IM-9 cell line, the predominance of mature PKC beta II message as well as that of a larger message spliced to PKC beta II provided evidence that the differential expression of PKC beta II was regulated at the level of mRNA splicing. In the BJA-B cell line, equal amounts of mature PKC beta I and beta II message and the absence of the larger message suggested that the splicing of the PKC beta gene product can be regulated to produce altered ratios of PKC beta I and beta II. Implications of these studies on the differential expression of PKC isoenzymes and their roles in biology are discussed.
Blobe
G C
GC
Department of Medicine, Duke University Medical Center, Durham, North Carolina 27710.
Khan
W A
WA
Halpern
A E
AE
Obeid
L M
LM
Hannun
Y A
YA
D017398:Alternative Splicing; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D011493:Protein Kinase C / Q000235:genetics*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D012260:Ribonucleases / Q000378:metabolism
D007527:Isoenzymes; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D011493:Protein Kinase C; D012260:Ribonucleases