Molecular cloning of platelet factor XI, an alternative splicing product of the plasma factor XI gene.
Platelet factor XI is associated with the platelet plasma membrane and has an apparent Mr (220,000 nonreduced, 55,000 reduced) different from that of plasma factor XI. However, the site of synthesis and the nature of platelet factor XI are not known. Using reverse transcriptase polymerase chain reaction, 12 out of 13 exons (all except exon V) coding for mature plasma factor XI were amplified from human platelet mRNA. The sequence of each of these exons was identical to that of plasma factor XI. In situ amplification and hybridization of factor XI mRNA was positive for exon III and negative for exon V in platelets and negative for both exons in other blood cells. By Northern hybridization, a factor XI mRNA transcript of approximately 1.9 kilobases was detected in megakaryocytic cells, and one of approximately 2.1 kilobases was detected in liver cells. Factor XI cDNA was cloned from a megakaryocyte library and sequenced. Exon V was absent, and the splicing of exon IV to exon VI maintained the open reading frame without alteration of the amino acid sequence except for the deletion of amino acids Ala91-Arg144 within the amino-terminal portion of the Apple 2 domain. Thus, platelet factor XI is an alternative splicing product of the factor XI gene, localized to platelets and megakaryocytes but absent from other blood cells.
Hsu
T C
TC
Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.
Shore
S K
SK
Seshsmma
T
T
Bagasra
O
O
Walsh
P N
PN
D017398:Alternative Splicing; D001483:Base Sequence; D001792:Blood Platelets / Q000378:metabolism*; D015152:Blotting, Northern; D003001:Cloning, Molecular; D015342:DNA Probes; D018076:DNA, Complementary; D005091:Exons; D005172:Factor XI / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D018076:DNA, Complementary; D012333:RNA, Messenger; D005172:Factor XI; D012194:RNA-Directed DNA Polymerase