Genomic organization and differential splicing of the mouse and human Pcyt2 genes.
CTP: ethanolaminephosphate cytidylyltransferase (Pcyt2) is an important regulatory enzyme in phosphatidylethanolamine and plasmalogen biosynthesis. We cloned the mouse gene mPcyt2 and established its relationship with the human homolog PCYT2. The two genes share similar size and contain two conserved catalytic domains but exhibit different exon/intron organization. An internal region could be alternatively spliced producing a longer mouse transcript, mPcyt2 alpha, and a shorter human transcript, PCYT2 beta. The spliced region is entirely made from mPcyt2 Exon 7 and encodes the peptide PPHPTPAGDTLSSEVSSQ, located upstream of the second catalytic motif HIGH. Mouse and human proteins also differ in amino acid composition at the C-terminus due to an additional splicing between Exons 13 and 14 in PCYT2. The 5' RACE analyses and subsequent cloning of the promoter regions demonstrated that the mPcyt2 and PCYT2 promoters are located immediately upstream of the first exon. There is no sequence homology between the two promoters but they are both TATA-less, have conserved CAAT boxes at a matching distance (-85/-70 bp) from the transcription start site and contain cis-elements for transcription factors of the CAAT, Sp1 and NF1 family, all in accordance with ubiquitous expression of both genes. The mPcyt2 gene is highly expressed in liver, brain, adipose tissues, heart, skeletal muscle, spleen, lungs and kidney. In THP-1 and U937 cells, PCYT2 expression could vary with the stage of cell differentiation. Luciferase reporter analyses show that the Pcyt2 and PCYT2 promoters are strong promoters similar to other ubiquitous promoters, such as those of Pcyt1 and SV-40.
Poloumienko
Arkadi
A
Department of Human Biology and Nutritional Sciences, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Coté
Atina
A
Quee
Angela Tie Ten
AT
Zhu
Lin
L
Bakovic
Marica
M
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005091:Exons; D005796:Genes / Q000235:genetics; D006801:Humans; D007438:Introns; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D012316:RNA Nucleotidyltransferases; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D024363:Transcription Initiation Site; D020298:U937 Cells
D011993:Recombinant Fusion Proteins; D004247:DNA; D008156:Luciferases; D009713:Nucleotidyltransferases; D012316:RNA Nucleotidyltransferases; C020126:Ethanolamine-phosphate cytidylyltransferase