Cloning and characterization of a novel caprine genomic repetitive element that hybridizes with papillomavirus DNA.
A goat genomic library was screened by Southern blot hybridization at reduced stringency with a bovine papillomavirus type 5 (BPV 5) DNA probe in order to identify potential cellular and viral sequences related to the papillomavirus genome. A recombinant clone with an 8.5 kb genomic insert was found to contain a 1.3 kb PstI subfragment (designated as P1-1) that hybridized with the DNA of BPV 5, two murine papillomaviruses and human papillomavirus types 5 and 8, but not with DNA from another eight human and bovine papillomavirus types. Southern blot hybridization of the goat P1-1 DNA probe was restricted to a single 1.0 kb subfragment within the E1 open reading frame (ORF) of BPV 5 but produced multiple bands ranging between 1.0 and 9.0 kb when hybridized under stringent conditions with PstI-digested DNA obtained from different goat tissues. The genomic sequence of P1-1 has direct repeats of 10 and 13 nucleotides flanking 153 nucleotides, and 889 nucleotides of sequence, respectively, and an inverted repeat sequence of 11 nucleotides flanking a major ORF potentially coding for 244 residues. Potential splice acceptor and donor sites capable of joining with upstream and downstream exons are present within the major ORF. Sequence similarity between P1-1 and BPV 5 DNA at the nucleotide and amino acid level was limited to a stretch of 58 nucleotides which includes an oligopurine/pyrimidine tract. This region of similarity contains a predicted glutamic acid-rich domain. The P1-1 sequence is a novel repetitive element within the goat genome that is unrelated in sequence to papillomavirus DNA and to genomic sequences of mouse and man.
Kulski
J K
JK
Centre for Molecular Immunology and Instrumentation, Department of Microbiology, The University of Western Australia, Nedlands, Australia. jkulski@cyllene.uwa.edu.au
Ward
B K
BK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D004247:DNA / Q000737:chemistry; D015344:DNA Probes, HPV; D004273:DNA, Neoplasm / Q000737:chemistry; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000737:chemistry*; D006041:Goats / Q000235:genetics*; D006801:Humans; D008221:Lymphoid Tissue / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D017385:Sequence Homology; D013945:Thymoma / Q000235:genetics; D013953:Thymus Neoplasms / Q000235:genetics
D015344:DNA Probes, HPV; D004273:DNA, Neoplasm; D004274:DNA, Recombinant; D004279:DNA, Viral; D004247:DNA