Deletion mutants that alter differential RNA processing in the E3 complex transcription unit of adenovirus

Bhat, Bheem M.; Brady, Helen; Pursley, Michael; Wold, William S.M.

doi:10.1016/0022-2836(86)90240-8

Cited by 30 publications

(19 citation statements)

References 39 publications

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“…gpl9K is coded by mRNAs a and b, and 14.7K protein is coded by mRNA h (30). AdS, rec700, and Ad2 make comparable levels of these mRNAs (2). As expected, these viruses make comparable levels of gpl9K and comparable levels of 14.7K protein.…”

Section: Resultssupporting

confidence: 69%

Identification and gene mapping of a 14,700-molecular-weight protein encoded by region E3 of group C adenoviruses

Tollefson

Wold

1988

J Virol

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Early region E3 of adenovirus type 5 should encode at least nine proteins as judged by the DNA sequence and the spliced structures of the known mRNAs. Only two E3 proteins have been proved to exist, a glycoprotein (gpl9K) and an 11,600-molecular-weight protein (11.6K protein). Here we describe an abundant 14.7K protein coded by a gene in the extreme 3' portion of E3. To identify this 14.7K protein, we constructed a bacterial vector which synthesized a TrpE-14.7K fusion protein, then we prepared antiserum against the fusion protein. This antiserum immunoprecipitated the 14.7K protein from cells infected with adenovirus types 5 and 2, as well as with a variety of E3 deletion mutants. Synthesis of the 14.7K protein correlated precisely with the presence or absence of the 14.7K gene and with the synthesis of the mRNA (mRNA h) which encodes the 14.7K protein. The 14.7K protein appeared as a triplet on immunoprecipitation gels and Western blots (immunoblots).

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Section: Resultssupporting

confidence: 69%

Identification and gene mapping of a 14,700-molecular-weight protein encoded by region E3 of group C adenoviruses

Tollefson

Wold

1988

J Virol

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“…Interestingly, however, there were demonstrable differences in the levels of CD expressed, as measured by the conversion of substrate, . 22 This subtle difference between the serotypes may imply some unknown regulatory elements in the 3Ј region of the Ad5 gp19 K gene. In ONYX-301 and -302, the engineered PmeI site is located within the 120 bp sequence unique to Ad5; however, in ONYX-303 and -304, the endogenous MunI site used for the 3Ј transgene insertion site is located downstream of this sequence, removing all of the Ad5 unique 120 bp sequence.…”

Section: Resultsmentioning

confidence: 99%

Gene delivery from the E3 region of replicating human adenovirus: evaluation of the 6.7 K/gp19 K region

Hawkins

Johnson²,

Bauzon³

et al. 2001

Gene Ther

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“…In certain differentially spliced genes, additional elements located upstream of the branch point (36, 58) or downstream of the 5' splice site (7) have been shown to influence recognition of the neighboring exon. In addition, splicing efficiency in a number of systems is influenced by exon sequences (5,6,11,16,18,20,22,23,28,29,36,37,40,51,59,(63)(64)(65)72). Thus, a region encompassing shortly upstream of the branch point to shortly downstream of the 5' splice site usually contains all of the sequences necessary for recognition of an exon.…”

mentioning

confidence: 99%

In vivo recognition of a vertebrate mini-exon as an exon-intron-exon unit.

Sterner¹,

Berget²

1993

Mol. Cell. Biol.

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Very small vertebrate exons are problematic for RNA splicing because of the proximity of their 3 ' and 5' splice sites. In this study, we investigated the recognition of a constitutive 7-nucleotide mini-exon from the troponin I gene that resides quite close to the adjacent upstream exon. The mini-exon failed to be included in spliced RNA when placed in a heterologous gene unless accompanied by the upstream exon. The requirement for the upstream exon disappeared when the mini-exon was internally expanded, suggesting that the splice sites bordering the mini-exon are compatible with those of other constitutive vertebrate exons and that the small size of the exon impaired inclusion. Mutation of the 5' splice site of the natural upstream exon did not result in either exon skipping or activation of a cryptic 5' splice site, the normal vertebrate phenotypes for such mutants. Instead, a spliced RNA accumulated that still contained the upstream intron. In vitro, the mini-exon failed to assemble into spliceosome complexes unless either internally expanded or accompanied by the upstream exon. Thus, impaired usage of the mini-exon in vivo was accompanied by impaired recognition in vitro, and recognition of the mini-exon was facilitated by the presence of the upstream exon in vivo and in vitro.Cumulatively, the atypical in vivo and in vitro properties of the troponin exons suggest a mechanism for the recognition of this mini-exon in which initial recognition of an exon-intron-exon unit is followed by subsequent recognition of the intron.All intron-containing pre-mRNAs contain specific conserved sequence elements at the 5' and 3' splice sites that are essential for accurate and efficient splicing. In vertebrates, the branch point, pyrimidine tract, 3' splice site, and 5' splice site are important determinants of splicing efficiency (recently reviewed in reference 27). Although each can diverge from consensus, the degree to which each matches the consensus is related to the efficiency of splicing of the adjacent exon (19,21,25,35,44,48,50,52,71,(73)(74)(75)(76). In certain differentially spliced genes, additional elements located upstream of the branch point (36, 58) or downstream of the 5' splice site (7) have been shown to influence recognition of the neighboring exon. In addition, splicing efficiency in a number of systems is influenced by exon sequences (5,6,11,16,18,20,22,23,28,29,36,37,40,51,59,(63)(64)(65)72). Thus, a region encompassing shortly upstream of the branch point to shortly downstream of the 5' splice site usually contains all of the sequences necessary for recognition of an exon. We have recently suggested that the exon is the unit of splice site recognition in vertebrates (53). This perspective suggests that successful exon recognition occurs when the balance of all of these intron and exon elements is sufficient to support the definition process.The length of the exon is also an important parameter in its recognition. The average size of vertebrate internal exons is 137 nucleotides (30). Experiments man...

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Deletion mutants that alter differential RNA processing in the E3 complex transcription unit of adenovirus

Cited by 30 publications

References 39 publications

Identification and gene mapping of a 14,700-molecular-weight protein encoded by region E3 of group C adenoviruses

Identification and gene mapping of a 14,700-molecular-weight protein encoded by region E3 of group C adenoviruses

Gene delivery from the E3 region of replicating human adenovirus: evaluation of the 6.7 K/gp19 K region

In vivo recognition of a vertebrate mini-exon as an exon-intron-exon unit.

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