Adenovirus types 2 and 5 functions elicit replication and late expression of adenovirus type 12 DNA in hamster cells

Abstract

Cells and virus. The following cell lines were propagated in Dulbecco modified Eagle medium (3) supplemented with 7 to 10% fetal bovine serum: BHK-21 cells, subline B3 (41), and the Ad2-transformed hamster cell lines HE1, HE3, HE5, and HE7 (7), as well as human HeLa cells (ATCC, CCL 2). Human KB cells (ATCC, CCL 17) were grown in Eagle medium containing 10% newborn calf serum. The AdStransformed cell line BHK297-131 (47), a gift of J. Sussenbach, Utrecht, The Netherlands, was grown in the Dulbecco modified Eagle medium containing 7% fetal calf serum. Cell lines HEl, HE3, HES, and HE7 were gifts of A

Late transcripts of adenovirus type 12 DNA are not translated in hamster cells expressing the E1 region of adenovirus type 5

“…In Adl2-infected BHK21 hamster cells, early but not late Adl2 mRNAs have been detected (21,24). Upon infection of the AdS-transformed hamster cell line BHK297-C131 with Adl1, however, late Adl2 RNAs are transcribed (15,16). In a comparative transcriptional analysis of dl12-infected human KB and Adl2-infected hamster BHK297-C131 cells, the production of poly(A)+ mRNAs has been studied by RNA transfer hybridization (Northern blotting) experiments (2,19) with DNA segments from the El (the EcoRI-C fragment), the L3 (the BarHI-F fragment), the E3 (the PstI-H fragment), the LL (the Pstl-P fragment), and the E4 (the BarnHI-E fragment) regions aS 32-P-labeled hybridization probes.…”

“…In Adl2-infected BHK297-C131 cells, early (El, E3, and E4) and late (L3, L5, pIVa, and pIX) regions of the reduced but clearly detectable amounts in comparison with the same regions transcribed in productively Adl2-infected human KB cells. In BHK21 cells devoid of integrated adenovirus genome fragments, Adl2 DNA replication and late gene transcription cannot be detected by the most sensitive techniques available (9,15,16). In spite of the presence of apparently nucleotide sequence-perfect, polyadenylated late Adl2 mRNAs, which are, at least in part, equipped with the authentic tripartite leader sequence, the synthesis of Adl2 fiber (or hexon) protein cannot be demonstrated in the complementing Adl2-infected BHK297-C131 system.…”

“…Hamster cells are permissive for human Ad2 and Ad5. In hamster cells carrying the El region of Ad5, a complementing system, Adl2 DNA can replicate, and late genes can be transcribed, but Adl2 virions are not produced (15,16). These findings raise the question of why the late Adl2-specific RNAs cannot be translated into the late virion structural proteins in the complemented system.…”

Epigenetic mechanisms in human adenovirus type 12 oncogenesis

“…In contrast, human Ad2 replicates in hamster cells [20,25]. Several attempts to salvage Ad12 DNA replication or steps beyond it in Syrian hamster cells have, at best, led to limited Ad12 DNA replication, even to late RNA synthesis in Ad5-transformed BHK297-C131 cells [26] but not to replication of Ad12 virions in substituting systems [21,22,24,[27][28][29]. Even after introducing amounts of Ad12 DNA comparable to that found in productively infected human cells, Ad12 DNA was still unable to overcome the barrier in Syrian hamster cells ( Fig.…”

Insufficient levels of NFIII and its low affinity for the origin of adenovirus type 12 (Ad12) DNA replication contribute to the abortive infection of BHK21 hamster cells by Ad12

“…In productively infected human cells, adenovirus replication proceeds in a well-regulated manner with early-gene expression being followed by viral DNA replication and the subsequent expression of the late viral functions. While the early E1 genes of Ad12 DNA are expressed in abortively infected hamster cells, there is a total block in Ad12 DNA replication (9,18,35) and the transcription of all late viral genes which is attributable to the failure of the Ad12 major late promoter (MLP) to function in hamster cells. A mitigator element in the downstream region of the MLP is likely responsible for the failure of the MLP to function in hamster cells (43,44).…”