An Imprinted PEG1/MEST Antisense Expressed Predominantly in Human Testis and in Mature Spermatozoa

• DOI: 10.1074/jbc.m200458200
• Publication Date: 2002-07-26T15:16:07Z
• Authors: Tao Li; Thanh H. Vu; Kok-Onn Lee; Youwen Yang; Chuyen V. Nguyen; Huy Q. Bui; Zhi-Lan Zeng; Binh T. Nguyen; Ji-Fan Hu; Susan K. Murphy; Randy L. Jirtle; Andrew R. Hoffman
• Keywords: Male; 0301 basic medicine; Base Sequence; Models, Genetic; Reverse Transcriptase Polymerase Chain Reaction; Molecular Sequence Data; Proteins; DNA Methylation; Oligonucleotides, Antisense; Blotting, Northern; Spermatozoa; Cell Line; Genomic Imprinting; 03 medical and health sciences; Protein Biosynthesis; Testis; Humans; CpG Islands; Tissue Distribution; RNA, Messenger; Luciferases; Alleles

PEG1 (or MEST) is an imprinted gene located on human chromosome 7q32 that is expressed predominantly from the paternal allele. In the mouse, Peg1/Mest is associated with embryonic growth and maternal behavior. Human PEG1 is transcribed from two promoters; the transcript from promoter P1 is derived from both parental alleles, and the transcript from P2 is exclusively from the paternal allele. We characterized the P1 and P2 transcripts in various normal and neoplastic tissues. In the normal tissues, PEG1 was transcribed from both promoters P1 and P2, whereas in six of eight neoplastic tissues, PEG1 was transcribed exclusively from promoter P1. Bisulfite sequencing demonstrated high levels of CpG methylation in the P2 region of DNA from a lung tumor. In the region between P1 and P2, we identified a novel transcript, PEG1-AS, in an antisense orientation to PEG1. PEG1-AS is a spliced transcript and was detected as a strong 2.4-kilobase band on a Northern blot. PEG1-AS and PEG1 P2-sense transcript were expressed exclusively from the paternal allele. Fragments of DNA from within the 1.5-kilobase region between PEG1-AS and the P2 exon were ligated to a pGL3 luciferase reporter vector and transfected into NCI H23 cells. This DNA exhibited strong promoter activity in both the sense and antisense directions, indicating that PEG1-AS and P2 exon share a common promoter region. Treatment of the transfected DNA fragments with CpG methylase abolished the promoter activity. Of interest, PEG1-AS was expressed predominantly in testis and in mature motile spermatozoa, indicating a possible role for this transcript in human sperm physiology and fertilization.