Cell type-specific proteins which interact with the 5' nontranslated region of hepatitis A virus RNA

Chang, Ki Ha; Brown, Edwin A.; Lemon, Stanley M.

doi:10.1128/jvi.67.11.6716-6725.1993

Cited by 61 publications

(46 citation statements)

References 37 publications

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“…Using UV cross-linking/label transfer assays, we show here that host proteins, in particular p38, p45, p57, and p84, specifically interact not only with the 3Ј-NTR of the HAV genome but also with sequences preceding the 3Ј-NTR and extending into the 3D pol -coding region. Interestingly, protein p38 was shown to interact also with the RNA sequences of the 5Ј-NTR and might be identical to a previously described IRES-binding protein (10).…”

mentioning

confidence: 56%

“…Preparation of cell extracts. At the same time postinfection, S-100 cell extracts were prepared from mock-and HAV-infected cells by the procedure described by Chang et al (10). Briefly, mock-or HAV-infected cells were removed from monolayers mechanically, collected by centrifugation at 150 to 200 ϫ g for 20 min at 4ЊC, and washed twice with phosphate-buffered saline.…”

Section: Methodsmentioning

confidence: 99%

“…UV cross-linking. UV cross-linking of RNA-protein complexes was performed as described by Chang et al (10). Briefly, cell extract (2 to 5 g of protein) was incubated for 20 min at 30ЊC with the radioactive RNA probe (0.5 ϫ 10 6 to 2 ϫ 10 6 cpm) in binding buffer (5 mM HEPES [pH 7.9], 25 mM KCl, 2 mM MgCl 2 , 1.75 mM ATP, 6 mM DTT, 0.05 mM phenylmethylsulfonyl fluoride, 0.05 mM EDTA, 5% glycerol) containing 30 g of tRNA (when indicated) in a total volume of 60 l. In competition experiments, the standard binding reaction was performed with unlabeled RNA in an approximately 50-fold molar excess before the radioactive RNA probe was added, and incubation continued at 30ЊC for further 20 min.…”

Section: Methodsmentioning

confidence: 99%

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RNA-protein interactions at the 3' end of the hepatitis A virus RNA

Kusov

Weitz

Dollenmeier

et al. 1996

J Virol

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The regulative cis-acting terminal RNA structures and the proteins involved in the amplification of the hepatitis A virus (HAV) genome are unknown. By UV cross-linking/label transfer experiments, we have analyzed sequences of the 3-nontranslated region (3-NTR) and preceding domains of the viral genome for their ability to interact with host proteins. A series of cDNA constructs were used to create genomic-and antigenomic-sense transcripts. The results show that the 3-NTR-poly(A) interacted with host cell proteins with molecular masses of 38, 45, 57, 84, and 110 kDa only weakly, compared with RNA structures also consisting of 3D-coding regions. Protein p38 was most efficiently labeled after interaction with secondarystructure elements located at the 3 end of the HAV RNA. p38 also interacted with a 5-terminal RNA probe. Optimal RNA binding was found to be dependent on the salt concentration. The specificity of the RNA-protein interaction was proven by competition assays. These data might indicate that a higher-order structure formed at the junction of the 3D pol -coding sequence and the 3-NTR of the HAV genome (putative RNA pseudoknot) significantly improves binding of host proteins and thus suggests that this structure might be essential for the formation of the replication complex initiating minus-strand RNA synthesis.

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mentioning

confidence: 56%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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RNA-protein interactions at the 3' end of the hepatitis A virus RNA

Kusov

Weitz

Dollenmeier

et al. 1996

J Virol

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“…In Figure 2A, RSW analysis was performed as previously described (Chang et al, 1993). In brief, exponentially growing T98G cells were collected by centrifugation (2000 r.p.m.)…”

Section: Fractionation and Rsw Analysismentioning

confidence: 99%

p97/DAP5 is a ribosome-associated factor that facilitates protein synthesis and cell proliferation by modulating the synthesis of cell cycle proteins

Lee

McCormick

2006

EMBO J

112

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p97 (also referred to as DAP5, NAT1 or eIF4G2) has been proposed to act as a repressor of protein synthesis. However, we found that p97 is abundantly expressed in proliferating cells and p97 is recruited to ribosomes following growth factor stimulation. We also report that p97 binds eIF2beta through its C-terminal domain and localizes to ribosome through its N-terminal MIF4G domain. When overexpressed, p97 increases reporter luciferase activity. In contrast, overexpression of the C-terminal two-thirds of eukaryotic initiation factor 4GI (eIF4GI), a region that shares significant homology with p97, or the N-terminal MIF4G domain of p97 markedly inhibits reporter activity, the rate of global translation and cell proliferation. Conversely, downregulation of p97 levels by RNA interference also decreases the rate of global translation and inhibits cell proliferation. This coincides with an increase in p27/Kip1 protein levels and a marked decrease in CDK2 kinase activity. Taken together, our results demonstrate that p97 is functionally different from the closely related C-terminal two-thirds of eIF4GI and it can positively promote protein synthesis and cell proliferation.

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“…In common to PV, HAV RNA was shown to have acquired mutations in its IRES element that enhance replication by facilitating cap-independent translation in a celltype-specific fashion. Interestingly, passage of HAV in different cell types engendered different sets of mutations; however, all adaptive events were clustered within the IRES [139][140][141].…”

Section: Translational Control and Viral Pathogenesismentioning

confidence: 99%

Translation initiation of viral mRNAs

López-Lastra

Ramdohr

Letelier

et al. 2010

Reviews in Medical Virology

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Viruses depend on cells for their replication but have evolved mechanisms to achieve this in an efficient and, in some instances, a cell-type-specific manner. The expression of viral proteins is frequently subject to translational control. The dominant target of such control is the initiation step of protein synthesis. Indeed, during the early stages of infection, viral mRNAs must compete with their host counterparts for the protein synthetic machinery, especially for the limited pool of eukaryotic translation initiation factors (eIFs) that mediate the recruitment of ribosomes to both viral and cellular mRNAs. To circumvent this competition viruses use diverse strategies so that ribosomes can be recruited selectively to viral mRNAs. In this review we focus on the initiation of protein synthesis and outline some of the strategies used by viruses to ensure efficient translation initiation of their mRNAs.

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Cell type-specific proteins which interact with the 5' nontranslated region of hepatitis A virus RNA

Cited by 61 publications

References 37 publications

RNA-protein interactions at the 3' end of the hepatitis A virus RNA

RNA-protein interactions at the 3' end of the hepatitis A virus RNA

p97/DAP5 is a ribosome-associated factor that facilitates protein synthesis and cell proliferation by modulating the synthesis of cell cycle proteins

Translation initiation of viral mRNAs

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