Polyomavirus large T-antigen binds the ‘pRb related’ protein p130 through sequences in conserved region 2

doi:10.1016/S0168-1702(96)01404-9

Virus Research

Volume 47, Issue 1, January 1997, Pages 85-90

https://doi.org/10.1016/S0168-1702(96)01404-9 Get rights and content

Abstract

The transforming potential of DNA tumor viruses derives mainly from the ability of their encoded oncogene products to interact with cellular proteins. Many of these viral oncoproteins share regions of sequence similarity, designated conserved region 1 and 2, which have been implicated in complex formation with pRb, the product of the retinoblastoma tumor suppressor gene, and related p107 and p130 species. It has now been shown that the E1A protein of adenovirus is able to bind to all three pRb-related proteins through sequences in conserved region 1 and 2. We have shown previously that polyomavirus large T-antigen also interacts with pRb and p107 in vitro. The pRb and p107 binding domains reside between residues 141–158, which include conserved region 2. In the present study, we demonstrate that polyomavirus large T-antigen also interacted with p130 in vitro through the same sequences in conserved region 2.

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Cited by (7)

Expression of RB2/p130 tumor-suppressor gene in AIDS-related non-Hodgkin’s lymphomas: Implications for disease pathogenesis
2002, Human Pathology
Citation Excerpt :
Although such a pattern of RB2/p130 and p107 expression has been found in other types of NHL,10,15 this was not the case in the present series of AIDS-related lymphomas. The absence of mutations in the RB2/p130 gene and the unusually high percentages of cells expressing pRb2/p130 in tumors with a high proliferative activity, such as AIDS-related lymphomas, may thus suggest a molecular mechanism similar to that observed in virus-linked oncogenesis.32-34 The results of our in vitro and in vivo binding assays indicate that Tat protein of HIV-1 is a viral protein that may physically interact with pRb2/p130.
In this study we examined 21 cases of AIDS-related lymphomas for genomic organization and expression of RB2/p130 oncosuppressor gene and compared the results with the proliferative features of these neoplasms. We found no mutations in the RB2/p130 gene and unusually high percentages of cells expressing nuclear pRb2/p130 in tumors with a high proliferative activity, such as AIDS-related lymphomas. These findings might suggest that a molecular mechanism usually observed in viral-linked oncogenesis could be involved. We performed in vitro and in vivo binding assays to investigate whether the human immunodeficiency virus (HIV) gene product Tat and Rb2/p130 could interact. The results of these assays revealed that the HIV-1 Tat protein binds specifically to pRb2/p130. This may result in the inactivation of its oncosuppressive properties and the induction of genes needed to proceed through the cell cycle including p107, cyclin A, and cyclin B. Using single-cell polymerase chain reaction (PCR) assay, we found HIV-1 DNA in the neoplastic cells of only 2 of the 21 cases examined, whereas PCR on whole tissue revealed HIV-1 DNA in all of the cases. Furthermore, a diffuse and nuclear stain was observed in tissue sections with anti-Tat monoclonal antibody. These findings are in accordance with the notion that soluble Tat protein could function as a biologically active extracellular protein released by infected cells and taken up readily by uninfected B cells. In conclusion, our results seem to suggest that pRb2/p130 oncosuppressor protein may be a target in the interaction between the HIV-1 gene products and host proteins. HUM PATHOL 33:723-731. Copyright 2002, Elsevier Science (USA). All rights reserved.
Genetic alterations of the retinoblastoma-related gene RB2/p130 identify different pathogenetic mechanisms in and among Burkitt's lymphoma subtypes
2000, American Journal of Pathology
Citation Excerpt :
Yet, a significant portion of E2F4 can exist as free E2F42 and/or can be bound to an as yet unidentified protein,43 thus driving cells to proliferation. In AIDS-related BL, the absence of mutations in the RB2/p130 gene and the unusually high levels of nuclear expression of both pRb2/p130 and p107 suggest that other mechanisms similar to the one observed in the virus-linked oncogenesis could be involved.44,45,47 All members of the retinoblastoma gene family share the ability to physically interact with certain oncoproteins of the DNA viruses.46
Alterations of cell cycle-associated genes probably contribute to the pathogenesis of Burkitt's Lymphoma (BL), in addition to c-myc translocation. Mutations disrupting the nuclear localization signal of the retinoblastoma-related gene RB2/p130 have been documented recently in BL cell lines and primary tumors. Given the importance of the RB2/p130 gene in controlling cell growth, mutations of this gene may result in uncontrolled cell proliferation. We tested the expression and genomic organization of the RB2/p130 gene in relation to the proliferative features of a series of BL samples collected from the endemic and sporadic regions, regardless of whether the samples were acquired immune deficiency syndrome (AIDS)-related. The expression of the Rb2/p130, p107, and cell proliferation-related proteins (cyclin A and B) was determined by immunohistochemistry. The structures of exons 19 through 22 of the RB2/p130 gene, encoding for the B domain and C terminus, were analyzed by polymerase chain reaction (PCR) analysis and single-strand conformation polymorphism (SSCP. technique. The direct PCR products were sequenced to identify the actual mutations. Our results suggest that BL is composed of a mixture of molecular types with distinct genetic and phenotypic patterns, probably resulting from different pathogenetic mechanisms. In endemic BL, the RB2/p130 gene is mutated in most of the cases, and the protein is restricted to the cytoplasm. In AIDS-related BL, high levels of nuclear expression of the wild-type pRb2/p130, p107, and cell proliferation-related proteins were detected. This finding is in line with the molecular mechanisms observed in virus-linked oncogenesis. Sporadic BLs were mainly characterized by the low nuclear values of the wild-type pRb2/p130 and, conversely, the high values of p107. The increased cell proliferation due to different alterations of cell growth control by Rb-related proteins may be the first step in lymphomagenesis, during which additional genetic changes, including missense mutations of c-myc, may subsequently occur.
Enhanced binding to origin DNA at low pH enables easy detection of polyomavirus large T antigen by gel mobility shift assay of unfixed complexes
1999, Journal of Virological Methods
Enhanced, stable binding by polyomavirus large T antigen to the viral DNA replication origin at pH 6 allowed the development of a gel mobility shift assay for the detection of large T antigen. Such assays were not possible at pH 7.6 without previous fixation, due to instability of the complexes. We demonstrated that the gel mobility shift assay at pH 6 is very sensitive, allowing the detection of as little as 5 ng large T antigen, and is highly specific for DNA containing G(A/G)GGC target sequences. This method was used to detect large T antigen in crude cell lysates from transformed yeast cell lines or nuclear extracts from infected insect cells. Large T antigen–DNA complexes remained at or near the loading well in 5% acrylamide or 1.5% agarose gels, indicating that these complexes are very large. Glycerol gradient analysis showed that protein–DNA complexes formed at pH 6 were massive, and that large T antigen also formed large complexes when incubated at low pH in the absence of DNA. These results show that pH has a major effect on binding of large T antigen to its multiple target sites in the viral origin of DNA replication, presumably by affecting protein–protein interactions that are important for the stability of large T antigen–DNA complexes.
Study on JV virus in patients with colon cancer type adenocarcinoma
2019, Asian Pacific Journal of Cancer Prevention
Global Analysis of Mouse Polyomavirus Infection Reveals Dynamic Regulation of Viral and Host Gene Expression and Promiscuous Viral RNA Editing
2015, PLoS Pathogens
Necdin, a p53-target gene, is an inhibitor of p53-mediated growth arrest
2012, PLoS ONE

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Short communicationPolyomavirus large T-antigen binds the ‘pRb related’ protein p130 through sequences in conserved region 2

Abstract

Cell

Cell

Cell

Cell

Cell

Cell

Cell

Cell

Cell

Cell

Functional synergy between DP-1 and E2F-1 in the cell cycle-regulating transcription factor DRTFI/E2F

EMBO J.

Functional interactions within adenovirus E1A protein complexes

Oncogene

Independent binding of the retinoblastoma protein and p107 to the transcription factor E2F

Nature

p130/pRb2 has growth suppressive properties similar to yet distinctive from those of retinoblastoma family members pRb and p107

Cancer Res.

Cell cycle-specific association of E2F with the p130 E1A-binding protein

Genes Dev.

Large T-antigens of many polyomaviruses are able to form complexes with the retinoblastoma protein

J. Virol.

The human papilloma virus-16 E7 oncoprotein is able to bind to the retinoblastoma gene product

Science

A point mutational analysis of human papillomavirus type 16 E7 protein

J. Virol.

Short communication
Polyomavirus large T-antigen binds the ‘pRb related’ protein p130 through sequences in conserved region 2