Localization of the E1 B proteins of adenovirus 5 in transformed cells, as revealed by interaction with monoclonal antibodies

Zantema, A.; Fransen, J.A.M.; Davis-Olivier, A; Ramaekers, Frans C.S.; Vooijs, G. P.; Deleys, Bob; Eb, A. J. Van Der

doi:10.1016/0042-6822(85)90421-0

Cited by 131 publications

(148 citation statements)

References 33 publications

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“…To elucidate this phenomenon, stably transformed rat cell lines AB120/AB115 expressing E1A plus E1B-55K-wt/E1A plus E1B-55K-NES were analysed for steady-state localization of E1B-55K and PML (Figures 1A and B). Consistent with the published results, E1B-55K-wt localizes in a large cytoplasmic accumulation in proximity to the nucleus (Figure 1Ad) (Zantema et al, 1985a, b), whereas PML exhibits a punctuate nuclear staining (Figure 1Ae) (Endter et al, 2005). However, on inhibition of the CRM1-mediated nuclear export by leptomycin B, E1B-55K-wt accumulates in several bright stained nuclear aggregates clearly colocalizing with endogenous PML in B70% of all cells examined (Figures 1Aj-Al).…”

Section: E1b-55k Interacts and Colocalizes With Endogenous Pml In Trasupporting

confidence: 89%

SUMO modification of E1B-55K oncoprotein regulates isoform-specific binding to the tumour suppressor protein PML

Wimmer

Schreiner

Everett³

et al. 2010

Oncogene

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The E1B-55K product from human adenovirus is a substrate of the small ubiquitin-related modifier (SUMO)-conjugation system. SUMOylation of E1B-55K is required to transform primary mammalian cells in cooperation with adenovirus E1A and to repress p53 tumour suppressor functions. The biochemical consequences of SUMO1 conjugation of 55K have so far remained elusive. Here, we report that E1B-55K physically interacts with different isoforms of the tumour suppressor protein promyelocytic leukaemia (PML). We show that E1B-55K binds to PML isoforms IV and V in a SUMO1-dependent and -independent manner. Interaction with PML-IV promotes the localization of 55K to PMLcontaining subnuclear structures (PML-NBs). In virusinfected cells, this process is negatively regulated by other viral proteins, indicating that binding to PML is controlled through reversible SUMOylation in a timely coordinated manner. These results together with earlier work are consistent with the idea that SUMOylation regulates targeting of E1B-55K to PML-NBs, known to control transcriptional regulation, tumour suppression, DNA repair and apoptosis. Furthermore, they suggest that SUMO1-dependent modulation of p53-dependent growth suppression through E1B-55K PML-IV interaction has a key role in adenovirus-mediated cell transformation.

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Section: E1b-55k Interacts and Colocalizes With Endogenous Pml In Trasupporting

confidence: 89%

SUMO modification of E1B-55K oncoprotein regulates isoform-specific binding to the tumour suppressor protein PML

Wimmer

Schreiner

Everett³

et al. 2010

Oncogene

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“…Thus, E1B-55K can clearly be considered a multifunctional protein. Since E1B-55K was also described as a cytoplasmic protein (Goodrum et al, 1996;Zantema et al, 1985) but obviously is involved in nuclear events like mRNA transport and transformation, we thoroughly examined the nucleo-cytoplasmic tra cking ability of E1B-55K. Here we show that the Ad5 E1B-55K protein shuttles independently from E4orf6 and cellular tra cking proteins like p53 or Mdm2.…”

Section: Introductionmentioning

confidence: 75%

“…Furthermore, it was shown recently that the adenovirus protein relieves growth constraints imposed on viral replication by the cell cycle Ornelles, 1997, 1999). Finally, it is interesting to note that Ad5 E1B-55K but not Ad12 E1B-54K colocalizes with the tumour suppressor proteins p53 and WT1 in a perinuclear body in transformed cells (Maheswaran et al, 1999;Zantema et al, 1985). Since Ad12 E1B-54K lacks the identi®ed NES (Figure 7) it is possible that the E1B-dependent cytoplasmic sequestration of both cellular proteins results from the ability of the Ad5 polypeptide to shuttle between the nucleus and the cytoplasm.…”

Section: Discussionmentioning

confidence: 99%

“…The Ad5 E1B-55K protein has been reported to localize to the cytoplasm, the nucleus or both depending on the cell line used and the presence or absence of other viral proteins (Goodrum et al, 1996;Rubenwolf et al, 1997;Zantema et al, 1985). In order to study E1B-55K localization and tra cking in live cells we expressed full length (referred to as E1B-GFP) and truncated E1B-55K proteins as hybrids with GFP ( Figure 1).…”

Section: E1b-55k Is An Active Shuttle Protein and Export Is Mediated mentioning

confidence: 99%

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The adenovirus type 5 E1B-55K oncoprotein is a highly active shuttle protein and shuttling is independent of E4orf6, p53 and Mdm2

et al. 2000

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The E1B-55K and E4orf6 oncoproteins of adenovirus type 5 are involved in the export of viral mRNAs. Previously, it was suggested that a complex composed of E1B-55K and E4orf6 serves as a nucleocytoplasmic transporter for viral mRNAs in which the E4orf6 protein directs both nuclear import and export. We now demonstrate that the E1B-55K protein itself shuttles e ciently in the absence of E4orf6. In addition, E1B-55K tra cking was independent of the de®ned shuttle proteins Mdm2 or p53, which interacts with E1B-55K. The identi®ed N-terminal E1B-55K leucine-rich nuclearexport signal (NES) conferred rapid nuclear export even in a heterologous system in contrast to the postulated E4orf6NES. Interestingly, although shuttling was blocked by inhibitors of the CRM1 mediated export pathway, E1B-55K inhibited neither the activity nor the tra cking of the retroviral shuttle proteins HIV-1 Rev and HTLV-1 Rex. In contrast, Rev or Rex blocked the nuclear export of E1B-55K, most likely by competing for essential export factors. Our results provide new insights into the regulation of the adenovirus mRNA export system and the processes of adenovirus mediated transformation. Oncogene (2000) 19, 850 ± 857.

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“…Thus, the reduction in p53 steady-state levels inversely correlated with the level of E4orf6 expression. Moreover, indirect immuno¯uorescence analyses revealed that the residual low levels of p53 in ABS1 cells were sequestered in cytoplasmic bodies (Zantema et al, 1985;Blair-Zajdel and Blair, 1988) which appeared to be much smaller relative to AB7 cells ( Figure 2b). As opposed to p53 the E4orf6 protein was localized in the nucleus of ABS1 cells (Figure 2c).…”

Section: Generation and Protein Analysis Of Transformed Brk Cellsmentioning

confidence: 97%

The adenovirus E4orf6 protein contributes to malignant transformation by antagonizing E1A-induced accumulation of the tumor suppressor protein p53

et al. 1999

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The adenovirus type 5 (Ad5) E4orf6 protein promotes focus formation of primary baby rat kidney (BRK) cells in cooperation with Ad5 E1 proteins. This activity is most likely related to the ability of the E4orf6 protein to bind to p53 and modulate its tumor suppressor functions. In this study we report that transformed BRK cells that stably express E4orf6 in addition to E1A and E1B (ABS cells) displayed multiple additional properties commonly associated with a high grade of oncogenic transformation compared to cells expressing only E1A and E1B (AB cells). These properties included morphological alterations, markedly enhanced growth rates and growth to much higher saturation densities. Following injection into nude mice ABS-derived tumors exhibited accelerated growth and, based on histopathological criteria, proofed to be much more malignant compared to tumors generated by AB cells. Interestingly, these highly transformed properties of ABS cells correlated with a dramatic reduction of p53 steady-state levels which inversely correlated with E4orf6 expression. From these results we conclude that expression of the Ad5 E4orf6 protein (i) confers additional transformed in vitro properties to primary rat cells expressing the Ad5 E1 proteins, and (ii) increases the tumorigenic and malignant potential of these cells in vivo. Our data suggest that the Ad5 E4orf6 protein enhances the intrinsic ability of E1-transformed rat cells to grow in a neoplastic state by completely inactivating p53 tumor suppressor function in combination with the E1A and E1B proteins.

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Localization of the E1 B proteins of adenovirus 5 in transformed cells, as revealed by interaction with monoclonal antibodies

Cited by 131 publications

References 33 publications

SUMO modification of E1B-55K oncoprotein regulates isoform-specific binding to the tumour suppressor protein PML

SUMO modification of E1B-55K oncoprotein regulates isoform-specific binding to the tumour suppressor protein PML

The adenovirus type 5 E1B-55K oncoprotein is a highly active shuttle protein and shuttling is independent of E4orf6, p53 and Mdm2

The adenovirus E4orf6 protein contributes to malignant transformation by antagonizing E1A-induced accumulation of the tumor suppressor protein p53

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