Identification of a molecular recognition feature in the E1A oncoprotein that binds the SUMO conjugase UBC9 and likely interferes with polySUMOylation

Yousef, Ahmed F.; Fonseca, Gregory J.; Pelka, Peter; Ablack, Jailal; Walsh, Charlotte Kirsten; Dick, Fred; Bazett-Jones, David P.; Shaw, Gary S.; Mymryk, Joe S.

doi:10.1038/onc.2010.226

Cited by 27 publications

(36 citation statements)

References 59 publications

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“…DREF was detected using a polyclonal anti-DREF antibody, GST was detected using a polyclonal anti-GST antibody, and input E1A was detected using the M73 antibody. E1A was previously found to alter the number and size of PML bodies in infected cells, we suspected that DREF was colocalized with PML bodies (31,33). Furthermore, SUMOylation is a key feature of many PML-associated proteins, and we observed altered SUMOylation of DREF (Fig.…”

Section: R (13s)supporting

confidence: 56%

“…E1A was previously reported to interact with UBC9, a SUMO conjugase (31,32). Interaction of E1A with UBC9 is required for alteration of PML body morphology in E1A-expressing cells (31), and this is abolished by the loss of E1A binding to UBC9 by the E1A T123H mutant.…”

Section: R (13s)mentioning

confidence: 97%

“…E1A is known to alter the subcellular distribution of proteins in order to alter their function (27), and we have previously observed altered PML body morphology upon expression of E1A in human cells (33) that was dependent on the ability of E1A to bind to UBC9 (31). Therefore, we next investigated the subcellular localization of DREF and whether E1A alters this distribution.…”

Section: R (13s)mentioning

confidence: 98%

“…Interaction of E1A with UBC9 is required for alteration of PML body morphology in E1A-expressing cells (31), and this is abolished by the loss of E1A binding to UBC9 by the E1A T123H mutant. We next determined whether different isoforms of E1A have differing effects on DREF SUMOylation and whether interaction with DREF or UBC9 was required for alteration of DREF SUMOylation by E1A.…”

Section: R (13s)mentioning

confidence: 99%

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Adenovirus E1A Targets the DREF Nuclear Factor To Regulate Virus Gene Expression, DNA Replication, and Growth

Radko

Koleva

James

et al. 2014

J Virol

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The adenovirus E1A gene is the first gene expressed upon viral infection. E1A remodels the cellular environment to maximize permissivity for viral replication. E1A is also the major transactivator of viral early gene expression and a coregulator of a large number of cellular genes. E1A carries out its functions predominantly by binding to cellular regulatory proteins and altering their activities. The unstructured nature of E1A enables it to bind to a large variety of cellular proteins and form new molecular complexes with novel functions. The C terminus of E1A is the least-characterized region of the protein, with few known binding partners. Here we report the identification of cellular factor DREF (ZBED1) as a novel and direct binding partner of E1A. Our studies identify a dual role for DREF in the viral life cycle. DREF contributes to activation of gene expression from all viral promoters early in infection. Unexpectedly, it also functions as a growth restriction factor for adenovirus as knockdown of DREF enhances virus growth and increases viral genome copy number late in the infection. We also identify DREF as a component of viral replication centers. E1A affects the subcellular distribution of DREF within PML bodies and enhances DREF SUMOylation. Our findings identify DREF as a novel E1A C terminus binding partner and provide evidence supporting a role for DREF in viral replication. IMPORTANCEThis work identifies the putative transcription factor DREF as a new target of the E1A oncoproteins of human adenovirus. DREF was found to primarily localize with PML nuclear bodies in uninfected cells and to relocalize into virus replication centers during infection. DREF was also found to be SUMOylated, and this was enhanced in the presence of E1A. Knockdown of DREF reduced the levels of viral transcripts detected at 20 h, but not at 40 h, postinfection, increased overall virus yield, and enhanced viral DNA replication. DREF was also found to localize to viral promoters during infection together with E1A. These results suggest that DREF contributes to activation of viral gene expression. However, like several other PML-associated proteins, DREF also appears to function as a growth restriction factor for adenovirus infection.T he interaction of the adenovirus early 1A (E1A) proteins with mammalian regulatory factors has been heavily exploited to elucidate the molecular basis by which they control cellular processes (1-5). Studying the interactions of E1A with new cellular targets provides an exciting opportunity to identify and dissect critical mechanisms controlling mammalian transcription, growth, and differentiation, as well as to identify novel viral regulatory mechanisms. The organization of E1A into short peptide motifs (MoRFs) (6) involved in protein interactions has significantly enriched our understanding of eukaryotic protein function. Identification and characterization of novel MoRFs within E1A allow their subsequent detection in other proteins, which suggests novel interactions leading to signifi...

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Section: R (13s)supporting

confidence: 56%

Section: R (13s)mentioning

confidence: 97%

Section: R (13s)mentioning

confidence: 98%

Section: R (13s)mentioning

confidence: 99%

See 2 more Smart Citations

Adenovirus E1A Targets the DREF Nuclear Factor To Regulate Virus Gene Expression, DNA Replication, and Growth

Radko

Koleva

James

et al. 2014

J Virol

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“…It has previously been shown that the viral early protein E1A represses SUMO modification of the retinoblastoma protein and interacts with the SUMO-conjugating enzyme Ubc9 to likely interfere with polySUMOylation (76,77). Therefore, we tested if E1A triggers reduced SUMO-2 modification of Sp100 (Fig.…”

Section: Sp100 Depletion Promotes Ad Progeny Production and Early Virmentioning

confidence: 99%

Sp100 Isoform-Specific Regulation of Human Adenovirus 5 Gene Expression

Berscheminski

Wimmer

Brun

et al. 2014

J Virol

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Promyelocytic leukemia nuclear bodies (PML-NBs) are nuclear structures that accumulate intrinsic host factors to restrict viral infections. To ensure viral replication, these must be limited by expression of viral early regulatory proteins that functionally inhibit PML-NB-associated antiviral effects. To benefit from the activating capabilities of Sp100A and simultaneously limit repression by Sp100B, -C, and -HMG, adenoviruses (Ads) employ several features to selectively and individually target these isoforms. Ads induce relocalization of Sp100B, -C, and -HMG from PML-NBs prior to association with viral replication centers. In contrast, Sp100A is kept at the PML tracks that surround the newly formed viral replication centers as designated sites of active transcription. We concluded that the host restriction factors Sp100B, -C, and -HMG are potentially inactivated by active displacement from these sites, whereas Sp100A is retained to amplify Ad gene expression. Ad-dependent loss of Sp100 SUMOylation is another crucial part of the virus repertoire to counteract intrinsic immunity by circumventing Sp100 association with HP1, therefore limiting chromatin condensation. We provide evidence that Ad selectively counteracts antiviral responses and, at the same time, benefits from PML-NB-associated components which support viral gene expression by actively recruiting them to PML track-like structures. Our findings provide insights into novel strategies for manipulating transcriptional regulation to either inactivate or amplify viral gene expression. IMPORTANCEWe describe an adenoviral evasion strategy that involves isoform-specific and active manipulation of the PML-associated restriction factor Sp100. Recently, we reported that the adenoviral transactivator E1A targets PML-II to efficiently activate viral transcription. In contrast, the PML-associated proteins Daxx and ATRX are inhibited by early viral factors. We show that this concept is more intricate and significant than originally believed, since adenoviruses apparently take advantage of specific PML-NBassociated proteins and simultaneously inhibit antiviral measures to maintain the viral infectious program. Specifically, we observed Ad-induced relocalization of the Sp100 isoforms B, C, and HMG from PML-NBs juxtaposed with viral replication centers. In contrast, Sp100A is retained at Ad-induced PML tracks that surround the newly formed viral replication centers, acting as designated sites of active transcription. The host restriction factors Sp100B, -C, and -HMG are potentially inactivated by active displacement from these sites, whereas Sp100A is retained to amplify Ad gene expression.

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SUMO Ubc9 enzyme as a viral target

2014

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Viruses alter specific host cell targets to counteract possible defense mechanisms aimed at eliminating infectivity and viral propagation. The SUMO conjugating enzyme Ubc9 functions as a hub for protein sumoylation, whilst also providing an interactive surface for sumoylated proteins through noncovalent interactions. The targeting of Ubc9 by viruses and viral proteins is thus highly beneficial for the disruption of both protein modification and protein-protein interaction mechanisms with which proteins increase their functional repertoire in cells. This review explores some of the clever mechanisms adopted by viruses to deregulate Ubc9, influence effector pathways and positively impact viral persistence consequently.

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Identification of a molecular recognition feature in the E1A oncoprotein that binds the SUMO conjugase UBC9 and likely interferes with polySUMOylation

Cited by 27 publications

References 59 publications

Adenovirus E1A Targets the DREF Nuclear Factor To Regulate Virus Gene Expression, DNA Replication, and Growth

Adenovirus E1A Targets the DREF Nuclear Factor To Regulate Virus Gene Expression, DNA Replication, and Growth

Sp100 Isoform-Specific Regulation of Human Adenovirus 5 Gene Expression

SUMO Ubc9 enzyme as a viral target

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