Characterization of cDNA clones for the alpha subunit of pyruvate dehydrogenase from Ascaris suum.

ABSTRACT

The pyruvate dehydrogenase complex of the adult parasitic nematode Ascaris suum functions in the reducing environment present in their anaerobic mitochondria. These organelles use fumarate and enoyl CoAs as terminal electron acceptors instead of oxygen. A lambda gt11 cDNA library was constructed from RNA isolated from adult ascarid muscle. Partial clones for the pyruvate dehydrogenase alpha subunit were isolated by screening the lambda gt11 library with a specific antiserum. Full-length clones (type I) were identified in a cDNA library prepared from RNA isolated from early embryos. During the hybridization screening, a second type of cDNA clone (type II) was identified. The nucleotide sequences of both clones are presented. The predicted amino acid sequences of the mature proteins are 91% identical to one another and about 55% identical to the predicted sequences of the alpha subunit of human pyruvate dehydrogenase. Northern blots were used to examine the expression of both mRNAs in various larval stages and in tissues of the adult. Type I sequences are found mainly in adult muscle. Type II sequences are abundant in third-stage larvae as well as in adult muscle.