A bicistronic Epstein-Barr virus mRNA encodes two nuclear proteins in latently infected, growth-transformed lymphocytes

Wang, F; Petti, Lisa M.; Braun, Daniel; Seung, Steven K.; Kieff, Elliott

doi:10.1128/jvi.61.4.945-954.1987

Cited by 113 publications

(42 citation statements)

References 71 publications

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“…It was suggested that in such cases, eucaryotic ribosomes are capable of terminationreinitiation reactions, thus allowing for the expression of downstream ORFs (see, e.g., references 20 and 21). Consistent with this, the polypeptide products encoded by the upstream minicistrons on several bicistronic mRNAs have been detected in vivo (7,10,34) and in vitro (4). The presence of upstream minicistrons generally acts to decrease the expression of the downstream major ORF (9,10,18,21,29), possibly because not all terminating ribosomes become competent for reinitiation.…”

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confidence: 63%

Mutational analysis of upstream AUG codons of poliovirus RNA

Pelletier

Flynn

Kaplan

et al. 1988

J Virol

121

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The 5' untranslated region of poliovirus type 2 Lansing RNA consists of 744 nucleotides containing seven AUG codons which are followed by in-frame termination codons, thus forming short open reading frames (ORFs). To determine the biological significance of these small ORFs, all of the upstream AUG codons were mutated to UUG. The point mutations were introduced into an infectious poliovirus cDNA clone, and RNA transcribed in vitro from the altered cDNA was transfected into HeLa cells to recover the virus. Mutation of AUG 7 resulted in a virus (called R2-5NC-14) with a small-plaque phenotype, whereas mutation of the other six AUG codons produced virus with a wild-type plaque morphology. To determine whether the small-plaque phenotype of R2-5NC-14 was due to altered translational efficiency of the viral mRNA, we constructed chimeric mRNAs containing the 5' noncoding region of poliovirus mRNA fused to the chloramphenicol acetyltransferase (CAT) coding sequence. mRNA containing a mutated AUG 7 codon showed decreased translational efficiency in vitro. The results indicate that the upstream ORFs of poliovirus RNA are not essential for viral replication and do not act as barriers to the translation of poliovirus mRNA. AUG 7 and flanking sequences may play a positive acting role in poliovirus RNA translation.

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confidence: 63%

Mutational analysis of upstream AUG codons of poliovirus RNA

Pelletier

Flynn

Kaplan

et al. 1988

J Virol

121

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“…Again, the broader antigenic response to EBV compared with KSHV is striking. The W1 exon of EBV BamHI-W encodes 22 amino acids that form part of the EBNA-LP protein [33,34]. The protein encoded by the entire EBV BamHI-W ORF (BWRF1) and the EBNA-LP protein were both printed on the array.…”

Section: Comparison Of Plasma From Hiv-positive Patients With Kshv-asmentioning

confidence: 99%

Comparison of Humoral Immune Responses to Epstein-Barr Virus and Kaposi’s Sarcoma–Associated Herpesvirus Using a Viral Proteome Microarray

Zheng

Wan²,

Cho

et al. 2011

The Journal of Infectious Diseases

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“…hsp72/hsc73 may mediate the previously observed interaction between EBNA-LP and the retinoblastoma protein or p53.The Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) plays a critical role in EBV-induced transformation (4,9). EBNA-LP mRNA is composed of repeating 22and 44-amino-acid-encoding exons (the W repeats) and two unique exons (Y1 and Y2) encoding the C-terminal 45 amino acids (15,21). EBV recombinants with a 45-amino-acid Cterminal truncated EBNA-LP due to a deletion of the Y1 and Y2 exons yielded 10-to 20-fold-fewer cell colonies in a fibroblast feeder layer soft agar assay than virus stocks derived from recombinant EBVs expressing an intact EBNA-LP (4).…”

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The Epstein-Barr virus nuclear antigen leader protein associates with hsp72/hsc73

Mannick

Xiao

Hemnes

et al. 1995

J Virol

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Epstein-Barr virus nuclear antigen leader protein (EBNA-LP) is important for primary B-lymphocyte growth transformation. We now demonstrate that the W repeat-encoded domain of EBNA-LP significantly associates with proteins of the heat shock protein 70 family (hsp72/hsc73). hsp72/hsc73 may mediate the previously observed interaction between EBNA-LP and the retinoblastoma protein or p53.The Epstein-Barr virus (EBV) nuclear antigen leader protein (EBNA-LP) plays a critical role in EBV-induced transformation (4,9). EBNA-LP mRNA is composed of repeating 22and 44-amino-acid-encoding exons (the W repeats) and two unique exons (Y1 and Y2) encoding the C-terminal 45 amino acids (15,21). EBV recombinants with a 45-amino-acid Cterminal truncated EBNA-LP due to a deletion of the Y1 and Y2 exons yielded 10-to 20-fold-fewer cell colonies in a fibroblast feeder layer soft agar assay than virus stocks derived from recombinant EBVs expressing an intact EBNA-LP (4). Recombinant viruses with a 45-amino-acid C-terminal deletion or stop codon truncation of EBNA-LP were even more markedly impaired in experiments using an endpoint dilution clonal transformation assay (9).The mechanism by which EBNA-LP affects transformation remains unknown although in vitro biochemical data have indicated a possible interaction with the retinoblastoma protein (RB) or p53. Glutathione-S-transferase (GST)-RB and GST-p53 bacterial fusion proteins can specifically bind a small fraction of EBNA-LP from cell lysates. Conversely, GST-EBNA-LP can bind approximately 1% of an RB bacterial fusion protein or an unknown fraction of p53 (19). To further elucidate EBNA-LP molecular interactions, we have immunoprecipitated EBNA-LP from cell extracts, sequenced an EBNA-LP-associated protein, and found that EBNA-LP associates with the heat shock protein 70 family (hsc73 and hsp72).The IB4 EBV-infected lymphoblastoid cell line (LCL) was labeled overnight with [ 35 S]methionine, and cellular proteins were extracted with an isotonic salt buffer (50 mM Tris [pH 8], 170 mM NaCl, 0.5% Nonidet P-40, 10 mg of apoprotinen per ml, 1 mM phenylmethylsulfonyl fluoride). The cellular proteins were then immunoprecipitated with an anti-EBNA-LP monoclonal antibody (JF186) (2) or an irrelevant anti-CD39 monoclonal antibody (AC2) (14), separated on an 8% polyacrylamide gel, and visualized with an autoradiograph. A 70-kDa protein was consistently present in the EBNA-LP monoclonal antibody immunoprecipitates but not in the irrelevant control monoclonal antibody immunoprecipitates, suggesting that this protein specifically associates with EBNA-LP (Fig. 1A). As can be seen in Fig. 1A, several other proteins were also immunoprecipitated by the EBNA-LP monoclonal antibody. However, these proteins were not evaluated because they were not consistently immunoprecipitated from EBNA-LP-positive cell

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A bicistronic Epstein-Barr virus mRNA encodes two nuclear proteins in latently infected, growth-transformed lymphocytes

Abstract: EBNA2 is a nuclear protein expressed in all cells latently infected with and growth transformed by Epstein-Barr virus (EBV) infection (K. Hennessy and E. Kieff, Science 227:1230-1240, 1985). The nucleotide sequence of the EBNA2 mRNA (J.

Cited by 113 publications

References 71 publications

Mutational analysis of upstream AUG codons of poliovirus RNA

Mutational analysis of upstream AUG codons of poliovirus RNA

Comparison of Humoral Immune Responses to Epstein-Barr Virus and Kaposi’s Sarcoma–Associated Herpesvirus Using a Viral Proteome Microarray

The Epstein-Barr virus nuclear antigen leader protein associates with hsp72/hsc73

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