The ribosomal protein L34 gene from the mosquito, Aedes albopictus: - exon-intron organization, copy number, and potential regulatory elements

We describe the structural analysis of genomic DNA encoding ribosomal prote in (rp) L34 from the mosquito, Aedes albopictus. Comparison of genomic DNA sequences encompassing approximately 8 kb with the rpL34 cDNA sequence show ed that the gene contains three exons and two introns, encoding a primary t ranscript with a deduced size of 6196 nucleotides from the transcription st art site to the polyadenylation site. Exon 1, which is not translated, meas ures only 45 bp, and is separated from Exon 2 by a 359 bp intron. Exon 2 me asures 78 bp, and contains the AUG translation initiation codon 14 nucleoti des downstream of its 5'-end. Downstream of Exon 2 is a 5270 bp intron, fol lowed by the remainder of the coding sequence in Exon 3, which measures 444 bp including the polyadenylation signal. We used a novel PCR-based procedu re to obtain 1.7 kb of DNA upstream of the rpL34 gene. Like the previously described Ae. albopictus rpL8 gene and various mammalian rp genes, the DNA immediately upstream of the rpL34 gene lacks the TATA box, and the rpL34 tr anscription initiation site is embedded in a characteristic polypyrimidine tract. The 5'-flanking DNA contained a number of cis-acting elements that p otentially interact with transcription factors characterized by basic domai ns, zinc-coordinating DNA binding domains, helix-turn-helix motifs, and bet a scaffold factors with minor groove contacts. Particularly striking was th e conservation of an AP-4 binding site within 100 nucleotides upstream of t he transcription initiation site in both Aal-rpL34 and Aal-rpL8 genes. Comp arison of Southern hybridization signals using probes from the 5' and 3'-en ds of the 5.3 kb second intron and the cDNA suggested that the Ae. albopict us rpL34 gene most likely occurs as a single expressed copy per haploid gen ome with restriction enzyme polymorphisms in the upstream flanking DNA and the likely presence of one or more pseudogenes. (C) 1999 Elsevier Science L td. All rights reserved.