Abstract
BackgroundThe human adenovirus type 5 early region 1A (E1A) gene encodes proteins that are potent regulators of transcription. E1A does not bind DNA directly, but is recruited to target promoters by the interaction with sequence specific DNA binding proteins. In mammalian systems, E1A has been shown to contain two regions that can independently induce transcription when fused to a heterologous DNA binding domain. When expressed in Saccharomyces cerevisiae, each of these regions of E1A also acts as a strong transcriptional activator. This allows yeast to be used as a model system to study mechanisms by which E1A stimulates transcription.ResultsUsing 81 mutant yeast strains, we have evaluated the effect of deleting components of the ADA, COMPASS, CSR, INO80, ISW1, NuA3, NuA4, Mediator, PAF, RSC, SAGA, SAS, SLIK, SWI/SNF and SWR1 transcriptional regulatory complexes on E1A dependent transcription. In addition, we examined the role of histone H2B ubiquitylation by Rad6/Bre1 on transcriptional activation.ConclusionOur analysis indicates that the two activation domains of E1A function via distinct mechanisms, identify new factors regulating E1A dependent transcription and suggest that yeast can serve as a valid model system for at least some aspects of E1A function.
Highlights
The human adenovirus type 5 early region 1A (E1A) gene encodes proteins that are potent regulators of transcription
These results show that the two activation domains of E1A function via distinct but overlapping mechanisms and suggest that yeast can serve as a valid model system for identifying new targets of E1A involved in transcriptional regulation
LexA DNA binding domain (DBD) fusions of E1A activate transcription in yeast E1A contains two independent regions that function as transcriptional activation domains when expressed as DBD fusions in mammalian cells
Summary
The human adenovirus type 5 early region 1A (E1A) gene encodes proteins that are potent regulators of transcription. Human adenovirus type 5 (HAdV-5) early region 1A (E1A) is the first viral gene expressed during infection and plays a critical role in transcriptional activation [1,2]. The primary E1A transcript is differentially spliced, yielding mRNAs encoding two major products of 289 residues (R) and 243R respectively (Figure 1A). These proteins share identical amino and carboxyl sequences and only differ by the presence of an additional 46 amino acids in the 289R protein [2,3]. An adjacent acidic region spanning residues 189–200, termed Auxiliary Region 1 (AR1), is essential for efficient transactivation of early viral promoters by E1A [12]
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