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Human adenovirus type 5 variants with sequence alterations flanking the E2A gene: Effects on E2 expression and DNA replication

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UNSPECIFIED (1996) Human adenovirus type 5 variants with sequence alterations flanking the E2A gene: Effects on E2 expression and DNA replication. VIRUS GENES, 12 (1). pp. 65-75. ISSN 0920-8569.

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Abstract

The human adenovirus type 5 (Ad5) E2 transcription unit is divided into a promoter-proximal region, E2A, and a distal region, E2B, each with its own polyadenylation site. Together these regions encode the three virus-derived proteins necessary for genome replication. Ad5 variants were produced that carried linker insertion mutations immediately 5' and/or 3' to the coding sequence for the E2A gene DNA binding protein (DBP). Two variants carrying solely a 5' lesion showed decreased usage of the adjacent 3' splice site, via which the DBP mRNA is produced, and an increased usage of the alternative downstream splice sites in the E2B region, wherein viral DNA polymerase and terminal protein precursor are encoded; these viruses showed somewhat reduced growth. A variant carrying a 3' lesion showed a marginal increase in DBP expression and slightly accelerated growth. When lesions 5' and 3' to the DBP coding sequence were combined in cis, the resulting virus was severely defective for growth and expressed E2B products to the virtual exclusion of E2A DBP. These data indicate that interactions must occur between the E2A 3' splice site and polyadenylation site before this region can be treated as an exon by the RNA processing machinery, and that a sequence alteration at the polyadenylation site that alone has only minor effects on the pattern of RNA processing can drastically affect terminal exon usage when placed in cis with a mutation that reduces splicing efficiency at the upstream 3' splice site. The data further indicate that, in vivo, Ad5 DNA replication is limited by prevailing DBP levels rather than by levels of polymerase or terminal protein precursor.

Item Type: Journal Article
Subjects: Q Science > QH Natural history > QH426 Genetics
Q Science > QR Microbiology > QR355 Virology
Journal or Publication Title: VIRUS GENES
Publisher: KLUWER ACADEMIC PUBL
ISSN: 0920-8569
Official Date: 1996
Dates:
DateEvent
1996UNSPECIFIED
Volume: 12
Number: 1
Number of Pages: 11
Page Range: pp. 65-75
Publication Status: Published

Data sourced from Thomson Reuters' Web of Knowledge

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