Enhanced expression of adenovirus transforming proteins

Gaynor, Richard B.; Tsukamoto, Ann; Montell, Craig; Berk, Arnold

doi:10.1128/jvi.44.1.276-285.1982

Cited by 88 publications

(47 citation statements)

References 41 publications

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“…When an adenovirus infection proceeds in the presence of an inhibitor of protein synthesis and the inhibitor is then removed, the synthesis of many of the early proteins (including those from EIA) is enhanced (Harter et al, 1976;Harter and Lewis, 1978), apparently because of an increase in the lifetime of early mRNA (Wilson et at., 1980). Gaynor et al (1982) have recently noted a similar phenomenon in cells infected for 24-48 h in the 850 Shuffling adenovirus promoters absence of viral DNA replication. If EIA protein synthesis is regulated translationally, these results argue that the regulation is confined to the late stage of infection, since conditions that maintain an infection at an early stage produce amplified levels of E1A proteins from amplified levels of E1A mRNAs.…”

Section: Synthesis Of Eia Proteinsmentioning

confidence: 63%

Shuffling adenovirus promoters: a viral recombinant with early region 1A under late transcriptional control.

Solnick¹

1983

The EMBO Journal

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Adenovirus recombinants were constructed in which a copy of the major late promoter was inserted either upstream or in place of the promoter for early region 1A (E1A). The ectopic late promoter directs a 20‐ to 100‐fold increase in the level of cytoplasmic RNA from E1A. The RNA is correctly processed at the normal splice and polyadenylation sites, and directs the in vitro synthesis of the full complement of E1A proteins. Furthermore, the novel transcripts are translated in vivo, since the recombinants are fully viable on HeLa cells notwithstanding the loss in one case of the intact E1A promoter.

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Section: Synthesis Of Eia Proteinsmentioning

confidence: 63%

Shuffling adenovirus promoters: a viral recombinant with early region 1A under late transcriptional control.

Solnick¹

1983

The EMBO Journal

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“…Bars: (B) 10 ,um, (F) 5 ,um, and (H) 5 have previously observed a nucleolar localization of hsp70 only in cells experiencing stress (e.g., after heat shock), it might be concluded that adenovirus infection simply represents yet another form of cellular stress. Further support for this idea follows from our observation that in 293 cells, which express ElA, E1B, and hsp7O, no obvious colocalization or coassociation of ElA and hsp70 was observed.…”

Section: Discussionmentioning

confidence: 99%

Differential distribution of the adenovirus E1A proteins and colocalization of E1A with the 70-kilodalton cellular heat shock protein in infected cells

White

Spector

Welch

1988

J Virol

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Five distinct localization patterns were observed for the adenovirus ElA proteins in the nuclei of infected HeLa cells: diffuse, reticular, nucleolar, punctate, and peripheral. The variable distribution of ElA was correlated with the time postinfection and the cell cycle stage of the host cell at the time of infection. All staining patterns, with the exception of peripheral ElA localization, were associated with the early phase of infection since only the diffuse, reticular, nucleolar, and punctate staining patterns were observed in the presence of hydroxyurea. Because the ElA proteins (12S and 13S) stimulate the expression of the cellular heat shock 70-kilodalton protein (hsp7O), we examined the intracellular distribution of hsp7O in the adenovirus-infected cells. Whereas hsp7O was predominantly cytoplasmic in the cells before infection, after adenovirus infection most of the protein was now found within the nucleus. Specifically, hsp7O was found within the nucleoli as well as exhibiting reticular, diffuse, and punctate nuclear staining patterns, analogous to those observed for the ElA proteins. Double-label indirect immunofluorescence of ElA and hsp7O in infected cells demonstrated a colocalization of these proteins in the nucleus. Translocation of hsp7O to the nucleus was dependent upon both adenovirus infection and expression of the ElA proteins. The localization of hsp7O was unaltered by infection with an ElA 9S cDNA virus which does not synthesize a functional ElA gene product. Moreover, the discrete nuclear localization patterns of ElA and the colocalization of ElA with hsp7O were not observed in adenovirus-transformed 293 cells which constitutively express ElA and ElB. ElA displayed exclusively diffuse nuclear staining in 293 cells; however, localization of ElA into the discrete nuclear patterns occurred after adenovirus infection of 293 cells. Immunoprecipitation of labeled infected-cell extracts with a monoclonal antibody directed against the ElA proteins resulted in precipitation of small amounts of hsp7O along with ElA. These data indicate that the adenovirus ElA proteins colocalize with, and possibly form a physical complex with, celHular hsp7O in infected cells. The relevance of this association, with respect to the function of these proteins during infection and the association of other oncoproteins with hsp7O, is discussed.

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“…RNA preparation and analysis To prepare RNAs, HeLa spinner cells were infected at a multiplicity of 20 p.f.u./cell and harvested either 5, 9 or 13 h later. RNA was prepared using the procedure of McGrogan et al (1979), and S1 endonuclease analysis using M13 single-stranded probes was essentially as described by Gaynor et al (1982). M13 ..1 I containing 8 M urea prepared in a Tris-borate buffer.…”

Section: Methodsmentioning

confidence: 99%

Dissection of overlapping functions within the adenovirus type 5 E1A gene.

Winberg¹,

Shenk²

1984

The EMBO Journal

144

113

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The adenovirus E1A gene encodes multiple, overlapping mRNAs whose products function both to regulate mRNA levels during the lytic cycle of the virus and to facilitate transformation of non‐permissive cells. To assign specific functions to the E1A gene products, two adenovirus type 5 variants have been constructed. Mutants dl347 and 348 carry cloned segments corresponding to the E1A 12 and 13S mRNAs, respectively, in place of the normal E1A gene. The variants produced the predicted E1A‐specific mRNAs and polypeptides. Both viruses grew efficiently in HeLa cells. Although the 13S mRNA products were more effective, the products of either mRNA species could stimulate the accumulation of mRNAs from additional transcription units. Both viruses could induce the formation of transformed foci in an established rat cell line. Neither virus could transform primary rat embryo cells at normal frequency, and the dl347 foci which were induced were incomplete or abortive transformants. Thus, functions encoded by both 12S and 13S mRNAs are required for efficient and complete transformation of primary rat cells.

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Enhanced expression of adenovirus transforming proteins

Cited by 88 publications

References 41 publications

Shuffling adenovirus promoters: a viral recombinant with early region 1A under late transcriptional control.

Shuffling adenovirus promoters: a viral recombinant with early region 1A under late transcriptional control.

Differential distribution of the adenovirus E1A proteins and colocalization of E1A with the 70-kilodalton cellular heat shock protein in infected cells

Dissection of overlapping functions within the adenovirus type 5 E1A gene.

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