Nuts and Bolts of Human Cytomegalovirus Lytic DNA Replication

Gs, Pari

doi:10.1007/978-3-540-77349-8_9

Cited by 40 publications

(56 citation statements)

References 56 publications

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“…Precisely how eIF4F contributes to viral DNA replication is currently unclear. Seven HCMV proteins are required for viral DNA replication in vitro (32). Perhaps one or more of these viral factors is dependent on eIF4F activity for their synthesis prior to viral DNA replication.…”

Section: Discussionmentioning

confidence: 99%

Differential Role for Host Translation Factors in Host and Viral Protein Synthesis during Human Cytomegalovirus Infection

Lenarcic

Ziehr

Leon

et al. 2014

J Virol

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The host eIF4F translation initiation complex plays a critical role the translation of capped mRNAs. Although human cytomegalovirus (HCMV) infection increases the abundance and activity of the host eIF4F complex, the requirement for eIF4F components in HCMV replication and mRNA translation has not been directly tested. In this study, we found that decreasing the abundance or activity of eIF4F from the start of infection inhibits HCMV replication. However, as infection progresses, viral mRNA translation and replication becomes increasingly resistant to eIF4F inhibition. During the late stage of infection the association of representative immediate-early, early, and late mRNAs with polysomes was not affected by eIF4F disruption. In contrast, eIF4F inhibition decreased the translation of representative host eIF4F-dependent mRNAs during the late stage of infection. A global analysis of the translation efficiency of HCMV mRNAs during the late stage of infection found that eIF4F disruption had a minimal impact on the association of HCMV mRNAs with polysomes but significantly diminished the translation efficiency of eIF4F-dependent host transcripts. Together, our data show that the translation of host eIF4F-dependent mRNAs remains dependent on eIF4F activity during HCMV infection. However, during the late stage of infection the translation efficiency of viral mRNAs does not correlate with the abundance or activity of the host eIF4F complex.

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Section: Discussionmentioning

confidence: 99%

Differential Role for Host Translation Factors in Host and Viral Protein Synthesis during Human Cytomegalovirus Infection

Lenarcic

Ziehr

Leon

et al. 2014

J Virol

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“…Although not widespread, there are several examples of RNA/DNA hybrids with biological significance, including their involvement in immunoglobin class switching (Tracy et al 2000), human cytomegalovirus virus lytic DNA replication (Pari 2008), and ColE1 plasmid replication (Santamaria et al 1998). The ease with which NMIA treatment can be combined with mass spectrometry should allow a more detailed analysis of these duplexes, either alone or in conjunction with their protein partners.…”

Section: Discussionmentioning

confidence: 99%

SHAMS: Combining chemical modification of RNA with mass spectrometry to examine polypurine tract-containing RNA/DNA hybrids

Turner¹,

Yi-Brunozzi²,

Brinson³

et al. 2009

RNA

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Selective 29-hydroxyl acylation analyzed by primer extension (SHAPE) has gained popularity as a facile method of examining RNA structure both in vitro and in vivo, exploiting accessibility of the ribose 29-OH to acylation by N-methylisatoic anhydride (NMIA) in unpaired or flexible configurations. Subsequent primer extension terminates at the site of chemical modification, and these products are fractionated by high-resolution gel electrophoresis. When applying SHAPE to investigate structural features associated with the wild-type and analog-substituted polypurine tract (PPT)-containing RNA/DNA hybrids, their size (20-25 base pairs) rendered primer extension impractical. As an alternative method of detection, we reasoned that chemical modification could be combined with tandem mass spectrometry, relying on the mass increment of RNA fragments containing the NMIA adduct (M r = 133 Da). Using this approach, we demonstrate both specific modification of the HIV-1 PPT RNA primer and variations in its acylation pattern induced by replacing template nucleotides with a non-hydrogen-bonding thymine isostere. Our selective 29-hydroxyl acylation analyzed by mass spectrometry strategy (SHAMS) should find utility when examining the structure of small RNA fragments or RNA/DNA hybrids where primer extension cannot be performed.

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“…Subsequently, HCMV replication sites are enlarged to form globular replication compartments that eventually fill the nucleus at late stage [19,29,30]. Viral proteins involved in HCMV DNA replication at least include UL44 (the DNA polymerase processivity factor), UL84 (likely a replication initiator interacting with the RNA/DNA hybrid within oriLyt) and the immediate-early 2 (IE2) protein [29,[31][32][33][34]. HCMV with IE2 gene deleted is not viable [35].…”

Section: Introductionmentioning

confidence: 99%

“…IE2 is a well studied transactivator essential for the induction of a cascade of HCMV lytic genes [36], some of which directly participate in viral DNA replication. Alone or in combination with UL84 that binds to a conserved RNA stem-loop within oriLyt [31,37,38], IE2 stimulates the expression of several genes within oriLyt essential for oriLyt activation [39]. In addition, Petrik et al [40] found that a recombinant HCMV carrying IE2 gene with an amino acid substitution at residue 548 from glutamine to arginine (Q548R) replicates slowly and loses the ability to arrest cell cycle.…”

Section: Introductionmentioning

confidence: 99%

Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2

Lee

et al. 2011

Cell Res

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Chromatin assembly factor 1 (CAF1) consisting of p150, p60 and p48 is known to assemble histones onto newly synthesized DNA and thus maintain the chromatin structure. Here, we show that CAF1 expression was induced in human cytomegalovirus (HCMV)-infected cells, concomitantly with global chromatin decondensation. This apparent conflict was thought to result, in part, from CAF1 mislocalization to compartments of HCMV DNA synthesis through binding of its largest subunit p150 to viral immediate-early protein 2 (IE2). p150 interaction with p60 and IE2 facilitated HCMV DNA synthesis. The IE2Q548R mutation, previously reported to result in impaired HCMV growth with unknown mechanism, disrupted IE2/p150 and IE2/histones association in our study. Moreover, IE2 interaction with histones partly depends on p150, and the HCMV-induced chromatin decondensation was reduced in cells ectopically expressing the p150 mutant defective in IE2 binding. These results not only indicate that CAF1 was hijacked by IE2 to facilitate the replication of the HCMV genome, suggesting chromatin assembly plays an important role in herpesviral DNA synthesis, but also provide a model of the virus-induced chromatin instability through CAF1.

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Nuts and Bolts of Human Cytomegalovirus Lytic DNA Replication

Cited by 40 publications

References 56 publications

Differential Role for Host Translation Factors in Host and Viral Protein Synthesis during Human Cytomegalovirus Infection

Differential Role for Host Translation Factors in Host and Viral Protein Synthesis during Human Cytomegalovirus Infection

SHAMS: Combining chemical modification of RNA with mass spectrometry to examine polypurine tract-containing RNA/DNA hybrids

Host-viral effects of chromatin assembly factor 1 interaction with HCMV IE2

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