Identification of a factor in HeLa cells specific for an upstream transcriptional control sequence of an EIA-inducible adenovirus promoter and its relative abundance in infected and uninfected cells.

Sivaraman, Lakshmi; Subramanian, Subhalakshmi; Thimmappaya, Bayar

doi:10.1073/pnas.83.16.5914

Cited by 111 publications

(90 citation statements)

References 39 publications

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“…If extracts were prepared at later times (20 hr postinfection), three-to fivefold lower levels of the factors binding to the BS2, BS3, and BS4 sites were seen (data not shown). This has been reported previously with respect to binding of ATF to the E2A promoter (Siva Raman et al 1986) and is probably due to sequestration of factors by the very high copy number of adenovirus DNA template generated by viral DNA replication. Interestingly, binding to the BS1 region was the same between uninfected and late-infected extracts.…”

Section: LIsupporting

confidence: 77%

“…Previous analysis of the E2A and E4 promoters indicated that similar factors might be interacting with specific regions of each promoter (Siva Raman et al 1986;Lee and Green 1987). Moreover, these same investigators also observed some slight competition with the E3 promoter, which we investigated further.…”

Section: Relationship Of the E3-binding Factors To Factors That Bind mentioning

confidence: 66%

“…Moreover, these same investigators also observed some slight competition with the E3 promoter, which we investigated further. Initially, we prepared a probe from the E2A promoter(-17 to -98) that contains a binding site for a factor E2A.EF, identified by Siva Raman et al (1986), and used this in gel-retention assays in the presence of E3 oligonucleotides. The DNA probe used contains a binding site for a second factor E2F, in addition to E2A.EF.…”

Section: Relationship Of the E3-binding Factors To Factors That Bind mentioning

confidence: 99%

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Identification of factors that interact with the E1A-inducible adenovirus E3 promoter.

Hurst

Jones²

1987

Genes Dev.

185

195

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We have investigated the E1A-inducible E3 promoter of adenovirus type 5 with respect to its ability to bind specific nuclear proteins. Four distinct nucleoprotein-binding sites were detected, located between positions -7 to -33, -44 to -68, -81 to -103, and -154 to -183, relative to the E3 cap site. These sites contain sequences previously shown to be functionally important for efficient E3 transcription. No major qualitative or quantitative differences were found in the binding pattern between nucleoprotein extracts prepared from uninfected or adenovirus-infected HeLa cells. Competition experiments suggest that the factors binding to the -154 to -183 and -81 to -103 sites are the previously identified nucleoproteins, NFI and AP1, respectively. The factor binding to the -44 to -68 site, which we term ATF, also interacts with other E1A-inducible promoters and is very similar and probably identical to the factor that binds to the cAMP-responsive element of somatostatin. We have purified this factor, which is a protein of 43 kD in size.

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Section: LIsupporting

confidence: 77%

Section: Relationship Of the E3-binding Factors To Factors That Bind mentioning

confidence: 66%

Section: Relationship Of the E3-binding Factors To Factors That Bind mentioning

confidence: 99%

See 1 more Smart Citation

Identification of factors that interact with the E1A-inducible adenovirus E3 promoter.

Hurst

Jones²

1987

Genes Dev.

185

195

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“…8) specific for adjacent DNA sequences between -69 and -33 is identified. Consistent with earlier reports (7,8), the levels of only this second factor, EIIF, increase considerably during viral infections.…”

supporting

confidence: 80%

“…Utilizing gel shift assays (4-6), we previously demonstrated that the uninfected HeLa cells contained a protein factor specific for the upstream transcriptional control element (between -82 and -66) of the EIA responsive Ad EIIA early (E) promoter (7). The levels of this factor did not change as a result of virus infection.…”

mentioning

confidence: 99%

Two promoter-specific host factors interact with adjacent sequences in an EIA-inducible adenovirus promoter.

Sivaraman¹,

Thimmappaya²

1987

Proc. Natl. Acad. Sci. U.S.A.

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We previously reported the identification of a host factor (EIIA-EF) specific for an upstream transcriptional control sequence (-82 to -66) of the EIA-inducible adenovirus EIIA early promoter. The levels of this factor remained unchanged after virus infection of human cells. Another study also identified a factor (EIIF) specific for this same promoter, but the activity of this second factor was shown to increase severalfold after virus infection. We now show that these dramatically different results, both based on gel shift assays on the same promoter, may be explained by variations in protocol details and actually identify two distinct factors. When synthetic DNA copolymers [poly(dI)-poly(dC) or poly(dI-dC)-poly(dI-dC)J are used as competitors in gel shift assays, a factor specific for DNA sequences between -82 and -66 can be identified, whereas when natural eukaryotic DNAs (salmon sperm or calf thymus) are used as competitors a different factor specific for DNA sequences between -69 and -33 can be identified. We have mapped the DNA-protein contact residues for the EIIF by analyzing a series of linker scan mutants in gel shift assays and methylation interference experiments. The EIIA-EF and EIIF bind to two distinct but adjacent sequences. Competition experiments indicate that these two activities are due to two different factors. Consistent with the earlier reports, the levels of one (EIIA-EF) do not change after virus infection of human cells, whereas the levels of the other (EIIF) are increased severalfold.Eukaryotic RNA polymerase TI promoters contain a complex array of cis-acting genetic elements that regulate basal, induced, and repressed transcription rates. These cis-acting regulatory sequences interact with a variety ofgeneral as well as gene-specific transcription factors. The interaction of the cis regulatory elements of promoters with the trans-acting promoter-specific DNA-binding proteins is important in control of tissue-specific, hormone-induced, viral-induced, and growth-related gene expression.Adenovirus (Ad) provides a useful model system to study the control of eukaryotic gene expression. In human cells infected with Ad type 2 or 5, a set offive early viral promoters are coordinately expressed (1). Efficient transcription of these early viral promoters and of several cellular promoters is dependent on the 32-kDa phosphoprotein encoded by the viral pre-early EIA gene. Recent negative results, including failure to detect either DNA-binding properties for the EIA protein (2) or sequence elements in the EIA responsive promoters recognized by the EIA gene product, have raised the possibility that the transcriptional activation by EIA may be mediated by promoter-specific host factors (for review, see ref.3). Viral infection of human cells may modify or increase the synthesis of host transcription factors. Alternatively, the transcription factors that are sequestered with the host promoters may upon infection be diverted to viral promoters by an as yet unidentified mechanism.Utilizing gel s...

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A cellular transcription factor E4F1 interacts with an E1a-inducible enhancer and mediates constitutive enhancer function in vitro.

Lee¹,

Green²

1987

The EMBO Journal

193

185

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Efficient transcription of the adenovirus Ela-inducible E4 gene is mediated by an Ela-dependent enhancer in vivo. In vitro, the enhancer functions constitutively in the absence of Ela, conferring high levels of transcription on the E4 and heterologous promoters. Binding of a cellular transcription factor, E4F1, to two sites within the E4 enhancer and to one other functionally important site located directly upstream of the E4 TATA box is required for transcriptional activity. The relationship between the enhancer and the TATA box proximal site is further demonstrated by the fact that the E4 enhancer can be functionally substituted by two copies of the TATA box proximal site. These and other results suggest that the E4 promoter may be comprised solely of multiple E4F1 binding sites and a TATA box. In addition to the E4 promoter, E4F1 interacts with other adenovirus early promoters and thus may be involved in the co-ordinate expression of Ela-inducible early viral genes.

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Identification of a factor in HeLa cells specific for an upstream transcriptional control sequence of an EIA-inducible adenovirus promoter and its relative abundance in infected and uninfected cells.

Cited by 111 publications

References 39 publications

Identification of factors that interact with the E1A-inducible adenovirus E3 promoter.

Identification of factors that interact with the E1A-inducible adenovirus E3 promoter.

Two promoter-specific host factors interact with adjacent sequences in an EIA-inducible adenovirus promoter.

A cellular transcription factor E4F1 interacts with an E1a-inducible enhancer and mediates constitutive enhancer function in vitro.

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