Evidence for two alternatively spliced forms of phospholipase C-beta2 in haematopoietic cells.
Alternatively spliced forms have been reported for several phospholipase C (PLC) isozymes, but not for PLC-beta2, the most abundant PLC-beta in platelets. PLC-beta2 cDNA cloned from the HL-60-cell cDNA library is 3543 bases long, coding for 1181 amino acids. Compared with the published sequence, a deletion of 45 nucleotides (2755-2799 nt, amino acids 864-878) was detected in platelet and leucocyte mRNA amplified by reverse transcription (RT) polymerase chain reaction (PCR) using primers corresponding to 1814-1838 nt (forward) and 3328-3352 nt (reverse). Amplification of genomic DNA using primers corresponding to 2575-2596 nt and 2864-2885 nt yielded a approximately 750 bp product; restriction analysis and sequencing revealed the 45-bp exon flanked by introns of 198 bp and 118 bp. Amplification of leucocyte and platelet cDNA using the same primers yielded products of approximately 310 nt and approximately 265 nt, with (PLC-beta2a) and without (PLC-beta2b) the 45-nt sequence. Thus, two alternatively spliced forms (1181 and 1166 amino acids) of PLC-beta2 are generated in haematopoietic cells. They differ in the carboxyl terminal sequence implicated in interaction of PLC-beta enzymes with Galphaq, particulate association and nuclear localization. We propose that the PLC-beta2 splice variants may be regulated differentially with distinct roles in signal transduction.
Mao
G F
GF
Sol Sherry Thrombosis Research Center and Departments of Medicine and Physiology, Temple University School of Medicine, Philadelphia, PA 19140, USA.
Kunapuli
S P
SP
Koneti Rao
A
A
D017398:Alternative Splicing / Q000235:genetics*; D001483:Base Sequence; D018076:DNA, Complementary / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D015398:Signal Transduction / Q000235:genetics; D010738:Type C Phospholipases / Q000235:genetics*
D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D010738:Type C Phospholipases