Cloning and characterization of the mouse cytochrome c oxidase subunit IV gene

Abstract

cDNA for mouse cytochrome c oxidase subunit IV (COXIV) was isolated by screening mouse liver and kidney cDNA libraries with a bovine COXIV cDNA probe. The 679-nucleotide nearly full length cDNA codes for a 22-amino acid presequence and a 147-amino acid mature protein which show 77 to 95% positional identity with the predicted sequences of human, bovine, and rat subunits. Screening of mouse genomic lambda EMBL3 library using the mouse cDNA probe yielded two overlapping clones. Restriction mapping and sequencing of the clones show that the mouse COXIV mRNA sequences are contained in five exons ranging from 58 to 236 base pairs, and four introns in a 7-kilobase region of the mouse genome. Southern blot analysis of restriction-digested genomic DNA indicates the presence of a single gene for COXIV in the mouse genome. Primer extension analysis using a synthetic 22-mer oligonucleotide, together with the 0.68-kilobase size of the mRNA shown by the Northern blot analysis, indicates that the major transcription start site of the COXIV gene is located 59 nucleotides upstream of the translation start site. The COXIV gene is highly GC rich and lacks TATA and CAAT elements in the immediate upstream region of the transcription start site. The putative promoter region, however, contains a number of GC boxes similar to those involved in the binding of Sp1 transcription factor. The unique features of the gene, as well as its characteristics common to other nuclear genes coding for different mitochondrial proteins, have been discussed.