Adenovirus Serotype Determines Association and Localization of the Large E1B Tumor Antigen with Cellular Tumor Antigen p53 in Transformed Cells

The human ERCC3 gene, which corrects specifically the nucleotide excision repair defect in human xeroderma pigmentosum group B and cross-complements the repair deficiency in rodent UV-sensitive mutants of group 3, encodes a presumed DNA helicase that is identical to the p89 subunit of the general transcription factor TFIIH/BTF2. To examine the significance of the postulated functional domains in ERCC3, we have introduced mutations in the ERCC3 cDNA by means of site-specific mutagenesis and have determined the repair capacity of each mutant to complement the UV-sensitive phenotype of rodent group 3 cells. A conservative substitution of arginine for the invariant lysine residue in the ATPase motif (helicase domain I), six deletion mutations in the other helicase domains, and a deletion in the potential helix-turn-helix DNA-binding motif fail to complement the ERCC3 excision repair defect of rodent group 3 mutants, which implies that the helicase domains as well as the potential DNA-binding motif are required for the repair function of ERCC3. Analysis of carboxy-terminal deletions suggests that the carboxy-terminal exon may comprise a distinct determinant for the DNA repair function. In addition, we show that a functional epitope-tagged version of ERCC3 accumulates in the nucleus. Deletion of the putative nuclear location signal impairs neither the nuclear location nor the repair function, indicating that other sequences may (also) be involved in translocation of ERCC3 to the nucleus.

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“…Immunoprecipitation and immunoblotting. Immunoprecipitation was performed essentially as described previously (56).…”

Section: Methodsmentioning

confidence: 99%

Mutational Analysis of ERCC3, Which Is Involved in DNA Repair and Transcription Initiation: Identification of Domains Essential for the DNA Repair Function

Westbroek

Jochemsen

et al. 1994

Molecular and Cellular Biology

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“…In light of our findings, we decided to evaluate whether ATM could be mediating the effects of Palbociclib onto p53 signalling axis by means of p53 Ser15 phosphorylation, a known target of ATM kinase activity implicated in DNA damage [33]. To this end, we took advantage of the high expression level of p53 in HT-29 and 293T cells, in which p53 is easily detected due to mutation or by the presence of viral proteins [34,35]. As it is shown in figure 7, p53 Ser15 phosphorylation was clearly increased in both cell lines after exposure to IR.…”

Section: P53 Is a Key Determinant Of Palbociclib-associated Radiosens...mentioning

confidence: 99%

P53 pathway is a major determinant in the radiosensitizing effect of Palbociclib: Implication in cancer therapy

et al. 2019

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Adenovirus early region 1A protein binds to mammalian SUG1-a regulatory component of the proteasome

et al. 1999

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Adenovirus early region 1A (Ad E1A) is a multifunctional protein which is essential for adenovirusmediated transformation and oncogenesis. Whilst E1A is generally considered to exert its in¯uence on recipient cells through regulation of transcription it also increases the level of cellular p53 by increasing the protein halflife. With this in view, we have investigated the relationship of Ad E1A to the proteasome, which is normally responsible for degradation of p53. Here we have shown that both Ad5 and Ad12 E1A 12S and 13S proteins can be co-immunoprecipitated with proteasomes and that the larger Ad12 E1A protein binds strongly to at least three components of the 26S but not 20S proteasome. One of these interacting species has been identi®ed as mammalian SUG1, a proteasome regulatory component which also plays a role in the cell as a mediator of transcription. In vitro assays have demonstrated a direct interaction between Ad12 E1A 13S protein and mouse SUG1. Following infection of human cells with Ad5 wt and Ad5 mutants with lesions in the E1A gene it has been shown that human SUG1 can be co-immunoprecipitated with full-length E1A and with E1A carrying a deletion in conserved region 1 which is the region considered to be responsible for increased expression of p53. We have concluded therefore that Ad E1A binds strongly to SUG1 but that this interaction is not responsible for inhibition of proteasome activity. This is consistent with the observation that puri®ed Ad12 E1A inhibits the activity of the puri®ed 20S but not 26S proteasomes. We have also demonstrated that SUG1 can be co-immunoprecipitated with SV40 T and therefore we suggest that this may represent a common interaction of transforming proteins of DNA tumour viruses.

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Adenovirus Serotype Determines Association and Localization of the Large E1B Tumor Antigen with Cellular Tumor Antigen p53 in Transformed Cells

Cited by 27 publications

References 36 publications

Mutational Analysis of ERCC3, Which Is Involved in DNA Repair and Transcription Initiation: Identification of Domains Essential for the DNA Repair Function

Mutational Analysis of ERCC3, Which Is Involved in DNA Repair and Transcription Initiation: Identification of Domains Essential for the DNA Repair Function

P53 pathway is a major determinant in the radiosensitizing effect of Palbociclib: Implication in cancer therapy

Adenovirus early region 1A protein binds to mammalian SUG1-a regulatory component of the proteasome

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