Characterization of a cDNA encoding a cytosolic peptidylprolyl cis-trans-isomerase from Blattella germanica

Abstract

Cyclophilins are an abundant and ubiquitous class of proteins first identified by their high affinity for the immunosuppressive drug cyclosporin A. Cyclophilins have peptidylprolyl cis/trans-isomerase activity in vitro, and thus may be involved in protein folding and trafficking in vivo. In this study, we report the cloning and characterization of a Blattella germanica cyclophilin cDNA. Analysis of this 846-bp cDNA reveals an open reading frame coding for a polypeptide of 164 amino acid residues with a molecular mass of 17934 Da. This B. germanica cyclophilin shares a central peptidylprolyl cis/trans-isomerase and a cyclosporin-A-binding domain with other cyclophilin sequences. The B. germanica cyclophilin amino acid sequence shares 83% identity with the cytosolic cyclophilin isoform from Drosophila melanogaster (Cyp-1). This identity suggests that B. germanica cyclophilin is a member of the cytosolic cyclophilin A (CyPA) family. From the alignment of cyclophilin sequences, we have found that 62 residues (positional identity of 40%) have remained invariant in eukaryotes for more than 1 billion years of divergence. We calculated a unit evolutionary period of 30.9 million years for the cytosolic isoform. Northern-blot analyses show that B. germanica CyPA mRNA is abundant, and present in all insect organs tested. The highest values for B. germanica cyclophilin mRNA tissue content were found in 6-day-old ovary, followed by brain, testis, and gut (15-30% the content of ovary). The muscle, fat body, and colleterial gland contained the lowest cyclophilin mRNA level (1-5% the content of ovary). There is a developmental pattern of gene expression affecting the embryo stages. These results suggest that this ubiquitously expressed B. germanica cyclophilin is subject to a differential regulation in tissues and during development. Southern-blot analysis of B. germanica DNA shows that only one copy of the CyPA gene is present per genome, whereas at least 20 genes or pseudogenes were detected in the mammalian genome.