Subgroup B adenovirus type 35 early region 3 mRNAs differ from those of the subgroup C adenoviruses

Adenovirus type 35 (Ad35) is a member of Ad subgroup B, DNA homology cluster B2. The B2 Ads are unique in that they are isolated most frequently from immunosupressed individuals such as AIDS patients and bone marrow transplant recipients and in that they have a tropism for the urinary tract. One region of the Ad genome which may influence serotype specific pathology is early region 3 (E3). E3 of subgroup C Ad2 and Ad5 has been shown to encode proteins which counteract the immune response to Ad infection. While a great deal is known about gene expression of the subgroup C Ad E3s, little is known about the E3 gene expression from the subgroup B Ads. Although some E3 open reading frames (ORFs) are shared between subgroups B and C, there are additional ORFs that appear in subgroup a. This paper demonstrates the results of an analysis of gene expression from the Ad35 E3 and describes differences in splicing and polyadenylation between the Ad35 and Ad2 E3s. RT-PCR, cDNA sequencing, RNase protection, 3' RACE, and Northern blotting techniques were utilized to identify, quantify, and determine the structure of six Ad35 E3 mRNAs predicted to encode at least seven proteins. A common intron that is removed during splicing of the subgroup C E3 mRNAs is not removed from Ad35 E3 mRNAs, and only one E3 polyadenylation signal is present in the Ad35 E3 while two polyadenylation signals are used in the formation of subgroup C E3 mRNAs. The quantity of individual mRNAs encoding homologous proteins for Ad35 and Ad2 also differ substantially, presumably because of the absence in Ad35 of cis-acting signals which have been shown to be important for regulation of Ad2 E3 pre-mRNA processing. Such information should contribute to an understanding of the role the E3 plays in determining subgroup B Ad pathogenesis in general and Ad35 pathogenesis in particular. (C) 1996 Academic Press, Inc.