Differentiation-specific expression from the bovine papillomavirus type 1 P2443 and late promoters.
The papillomavirus life cycle is tightly linked with keratinocyte differentiation in squamous epithelia. Vegetative viral DNA replication begins in the spinous layer, while synthesis of capsid proteins and virus maturation is restricted to the most differentiated or granular layer of the epithelium. In this study, in situ hybridization of bovine fibropapillomas was used to demonstrate that the activity of two promoters of bovine papillomavirus type 1 (BPV-1) is regulated in a differentiation-specific manner. In situ hybridization with a late promoter (PL)-specific oligonucleotide probe suggested that PL is dramatically upregulated in the granular layer of the fibropapilloma. Northern (RNA) blot analysis of RNA from BPV-1-infected fibropapillomas indicated that the three major BPV-1 late-region mRNAs were transcribed from PL. These RNAs include the previously described L1 (major capsid) mRNA as well as two larger mRNAs. The two larger mRNAs were characterized and shown to contain the L2 (minor capsid protein) open reading frame as well as the L1 open reading frame. In contrast to PL, the P2443 promoter was maximally active in basal keratinocytes and the fibroma. The major mRNA transcribed from P2443 is the putative E5 oncoprotein mRNA which is spliced between nucleotides 2505 and 3225. No signal was detected above the basal layer with use of a probe specific for this mRNA. The E5 oncoprotein has previously been localized by immunoperoxidase staining to the granular cell layer as well as the basal cell layer of the fibropapilloma (S. Burnett, N. Jareborg, and D. DiMaio, Proc. Natl. Acad. Sci. USA 89:5665-5669, 1992). These data suggest that E5 proteins in the basal cell and granular cell layers are not translated from the same mRNA.
Barksdale
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SK
Laboratory of Pathology, National Cancer Institute, Bethesda, Maryland 20892.
Baker
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CC
D000818:Animals; D001483:Base Sequence; D052688:Bovine papillomavirus 1 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002213:Capsid / Q000096:biosynthesis / Q000235:genetics; D002417:Cattle; D002418:Cattle Diseases; D002454:Cell Differentiation; D003001:Cloning, Molecular; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology; D005350:Fibroma / Q000382:microbiology / Q000473:pathology / Q000662:veterinary; D015967:Gene Expression Regulation, Viral; D017403:In Situ Hybridization; D015603:Keratinocytes / Q000166:cytology / Q000382:microbiology; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D010212:Papilloma / Q000382:microbiology / Q000473:pathology / Q000662:veterinary; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics
D004279:DNA, Viral; D004268:DNA-Binding Proteins; C051550:E2 protein, Bovine papillomavirus; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins