Abstract

Here we wish to focus attention on the role of DNA supercoiling in chromatin structure and gene expression. We have selected the powerful biological system of Saccharomyces cerevisiae to address the four following specific aims: (1) To elucidate the effect of DNA template linearization in vivo on transcription. -role of the position of the linearization relative to the polarity of a transcription unit -role of the position of the linearization relative to the positions of potential topological boundaries -minichromosomal versus chromosomal genes -role of telomere seed sequences flanking the site(s) of linearization (2) To delineate the mechanism(s) of nucleosome splitting -role of positive DNA supercoiling -role of torsional stress -role of RNA polymerase traversal -role of a helix tracking protein (3) To ascertain whether a template containing split nucleosomes is more efficiently transcribed compared to one containing whole nucleosomes (4) To determine whether DNA replication is required for the repair of split nucleosomes

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM022201-22
Application #
2518885
Study Section
Molecular Biology Study Section (MBY)
Project Start
1978-05-01
Project End
1999-08-31
Budget Start
1997-09-01
Budget End
1999-08-31
Support Year
22
Fiscal Year
1997
Total Cost
Indirect Cost

  Institution

Name
University of Texas Sw Medical Center Dallas
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390

  Publications

Sathyanarayana, U G; Freeman, L A; Lee, M S et al. (1999) RNA polymerase-specific nucleosome disruption by transcription in vivo. J Biol Chem 274:16431-6
Qiu, P; Kupfer, K C; Garrard, W T (1997) A method for genome comparisons and hybridization studies using known megabase-scale DNA sequences as a reference. Genomics 43:307-15
Liang, C P; Garrard, W T (1997) Template topology and transcription: chromatin templates relaxed by localized linearization are transcriptionally active in yeast. Mol Cell Biol 17:2825-34
Fishel, B R; Sperry, A O; Garrard, W T (1993) Yeast calmodulin and a conserved nuclear protein participate in the in vivo binding of a matrix association region. Proc Natl Acad Sci U S A 90:5623-7
Lee, M S; Garrard, W T (1992) Uncoupling gene activity from chromatin structure: promoter mutations can inactivate transcription of the yeast HSP82 gene without eliminating nucleosome-free regions. Proc Natl Acad Sci U S A 89:9166-70
Freeman, L A; Garrard, W T (1992) DNA supercoiling in chromatin structure and gene expression. Crit Rev Eukaryot Gene Expr 2:165-209
Whitehurst, C; Henney, H R; Max, E E et al. (1992) Nucleotide sequence of the intron of the germline human kappa immunoglobulin gene connecting the J and C regions reveals a matrix association region (MAR) next to the enhancer. Nucleic Acids Res 20:4929-30
Blasquez, V C; Hale, M A; Trevorrow, K W et al. (1992) Immunoglobulin kappa gene enhancers synergistically activate gene expression but independently determine chromatin structure. J Biol Chem 267:23888-93
Lee, M S; Garrard, W T (1991) Positive DNA supercoiling generates a chromatin conformation characteristic of highly active genes. Proc Natl Acad Sci U S A 88:9675-9
Lee, M S; Garrard, W T (1991) Transcription-induced nucleosome 'splitting': an underlying structure for DNase I sensitive chromatin. EMBO J 10:607-15

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