Deletion mutants which affect the nuclease-sensitive site in simian virus 40 chromatin.

Gerard, Robert D.; Woodworth-Gutai, M; Scott, Walter A.

doi:10.1128/mcb.2.7.782

Cited by 32 publications

(21 citation statements)

References 29 publications

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“…BSC-1 monkey cells were propagated as previously described (11). The nondefective mutant of SV40, in(Or)1411 (24), was grown from a stock provided by T. Shenk.…”

Section: Methodsmentioning

confidence: 99%

“…The nondefective mutant of SV40, in(Or)1411 (24), was grown from a stock provided by T. Shenk. Deletion mutants analyzed in this paper were isolated from in(Or)1411 as previously described (11), and their sequences were determined by the method of Maxam and Gilbert (19). Mutant ddl308 is a double-deletion mutant isolated from d145 and d142 (a 28-bp deletion within the ori site [11]) as described by Konig and Lai (18).…”

Section: Methodsmentioning

confidence: 99%

“…Deletion mutants analyzed in this paper were isolated from in(Or)1411 as previously described (11), and their sequences were determined by the method of Maxam and Gilbert (19). Mutant ddl308 is a double-deletion mutant isolated from d145 and d142 (a 28-bp deletion within the ori site [11]) as described by Konig and Lai (18). Mutant d1608 was isolated by targeting S1 endonuclease activity to the 163-bp segment of the in(Or)1411 genome between the BglI and BstNI sites by the D-loop procedure of Green and Tibbetts (12).…”

Section: Methodsmentioning

confidence: 99%

“…All essential information required for the hypersensitive site is contained within a 273-base-pair (bp) segment of DNA spanning the origin of replication (11,30 We have investigated the genetic elements responsible for the chromatin structure in this region by analyzing a number of additional mutants. No single segment is essential for nuclease sensitivity.…”

mentioning

confidence: 99%

“…All essential information required for the hypersensitive site is contained within a 273-base-pair (bp) segment of DNA spanning the origin of replication (11,30). Insertion of unrelated sequences between nucleotides 37 and 38 (SV numbering system of Buchman et al [4]) moves the pattern of nuclease cleavage sites in the late direction, indicating that sequences responsible for the positions of these sites lie on the side of nucleotide 37 which is distal from the origin of replication (15).…”

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confidence: 99%

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Role of specific simian virus 40 sequences in the nuclease-sensitive structure in viral chromatin.

Gerard¹,

Montelone²,

Walter³

et al. 1985

Mol. Cell. Biol.

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A nuclease-sensitive region forms in chromatin containing a 273-base-pair (bp) segment of simian virus 40 DNA encompassing the viral origin of replication and early and late promoters. We have saturated this region with short deletion mutations and compared the nuclease sensitivity of each mutated segment to that of an unaltered segment elsewhere in the partially duplicated mutant. Although no single DNA segment is required for the formation of a nuclease-sensitive region, a deletion mutation (d145) which disrupted both exact copies of the 21-bp repeats substantially reduced nuclease sensitivity. Deletion mutations limited to only one copy of the 21-bp repeats had little, if any, effect. A mutant (d1135) lacking all copies of the 21-and 72-bp repeats, while retaining the origin of replication and the TATA box, did not exhibit a nuclease-sensitive region. Mutants which showed reduced nuclease sensitivity had this effect throughout the nuclease-sensitive region, not just at the site of the deletion, indicating that although multiple determinants must be responsible for the nuclease-sensitive chromatin structure they do not function with complete independence. Mutant d19, which lacks the late portion of the 72-bp segment, showed reduced accessibility to BglI, even though the Bgll site is 146 bp away from the site of the deletion.Simian virus 40 (SV40)-infected cells contain SV40 DNA in a nucleoprotein structure with many similarities to cellular chromatin. The availability of an amplified, homogeneous DNA species in a chromatin-like structure has provided convenient material for the study of specific features of chromatin. Of particular interest is the short region adjacent to the viral origin of replication (ori site) which is hypersensitive to cleavage by endonucleases (23,26,27,29) and which appears as a gap in the nucleosome pattern when visualized in the electron microscope (16,22). This region contains the early and late gene promoters, the transcriptional enhancer sequences, and the viral origin of replication. The distinctive chromatin structure may be relevant to any or all of these functions.To identify which sequences are responsible for the nuclease-sensitive region, partially duplicated SV40 mutants have been studied. All essential information required for the hypersensitive site is contained within a 273-base-pair (bp) segment of DNA spanning the origin of replication (11,30 We have investigated the genetic elements responsible for the chromatin structure in this region by analyzing a number of additional mutants. No single segment is essential for nuclease sensitivity. However, overall accessibility to nuclease was dramatically decreased by a mutant affecting only the two exact copies of the 21-bp repeated sequences. Insertion of a foreign DNA segment into this region also substantially diminished nuclease sensitivity. By contrast, deletion of only one of the two exact copies of the 21-bp repeats had little effect on the pattern of nuclease cleavage.These results imply that the region of the genome e...

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“…BSC-1 monkey cells were propagated as previously described (11). The nondefective mutant of SV40, in(Or)1411 (24), was grown from a stock provided by T. Shenk.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Role of specific simian virus 40 sequences in the nuclease-sensitive structure in viral chromatin.

Gerard¹,

Montelone²,

Walter³

et al. 1985

Mol. Cell. Biol.

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SP1 and AP-1 Elements Direct Chromatin Remodeling in SV40 Chromosomes during the First 6 Hours of Infection

Milavetz¹

2002

Virology

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To identify the SV40 regulatory sequences responsible for the chromatin remodeling associated with early transcription, SV40 chromosomes containing potential remodeling sequences inserted adjacent to a reporter region were isolated at various times within the first 6 h of infection and analyzed by a combination of restriction endonuclease digestion and competitive PCR amplification. The sequences analyzed included the early domain, the enhancer, the late domain, the early phasing element, the AP-1 element, two tandem copies of the SP1 element, and the AP-4 element. From 30 min to 3 h postinfection only the enhancer, the AP-1 element, and the two tandem copies of the SP1 element caused a change in nuclease sensitivity consistent with chromatin remodeling. These results suggest that the changes in chromatin structure seen in the promoter during activation of early transcription are most likely a result of remodeling by the AP-1 and/or SP1.

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SV40 Chromatin Structure

Scott

1987

Molecular Aspects of Papovaviruses

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Deletion mutants which affect the nuclease-sensitive site in simian virus 40 chromatin.

Cited by 32 publications

References 29 publications

Role of specific simian virus 40 sequences in the nuclease-sensitive structure in viral chromatin.

Role of specific simian virus 40 sequences in the nuclease-sensitive structure in viral chromatin.

SP1 and AP-1 Elements Direct Chromatin Remodeling in SV40 Chromosomes during the First 6 Hours of Infection

SV40 Chromatin Structure

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