Abstract

Two overlapping clones containing the entire 684-nucleotide (nt) sequence encoding murine protein β-aspartate methyltransferase (EC 2.1.1.77) were isolated from a genomic library. Partial nt sequence analysis of the two clones revealed that the protein carboxyl methyltransferase (PCMT)-encoding sequence is distributed among seven exons, ranging from 32 to 339 bp in length, within 25 kb of genomic DNA. Three exons correspond to regions of primary structure which are strongly conserved among a number of eukaryotic and prokaryotic enzymes which utilize S-adenosylmethionine (AdoMet) and S-adenosylhomocysteine (AdoHcy). The 5'-flanking region of the PCMT-encoding gene ( PCMT) contains an 800-bp G+C-rich region with potential binding sites for transcription factor ETF, but lacks a TATA box and binding sites for other known transcription factors. Multiple PCMT mRNAs were detected on Northern blots of RNA extracted from murine brain, testis, liver and kidney. The overall abundance of PCMT mRNAs in each tissue paralleled the measured specific activity of the PCMT. Comparison of the genomic sequence information with the 3'-untranslated regions ( UTRs) of two cDNA clones from a murine testis library indicated that PCMT mRNA precursors undergo alternative splicing. The structure and widespread expression of PCMT are characteristics of vertebrate housekeeping genes.

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