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Printable Handouts
Navigable Slide Index
- Introduction
- Four basic stages of transcription
- Nucleosome organization: movie
- Outline of talk
- Nucleosome mapping: MNase ChIP-seq
- Peak shift by 73 bp to nucleosome center (1)
- Peak shift by 73 bp to nucleosome center (2)
- Browser map of nucleosome positions (1)
- Browser map of nucleosome positions (2)
- Nucleosomal arrays ordered by their length
- Canonical positioning of nucleosomes in yeast
- Nucleosomal context of regulatory elements
- Unbound transcription factor sites
- Fly nucleosomes compared to yeast
- The function of nucleosome organization
- RNA polymerase II pausing
- Paused vs. non paused RNA
- Distribution of nucleosomes that contact Pol II
- Position-specific histone H3 marks
- Position-specific histone modifications
- Regulatory proteins bind specific positions
- MNase digestion instead of sonication
- Reb1 interacts with the -1 nucleosome
- Rap1 interacts with the -1 nucleosome
- Chromatin regulator-nucleosome interactions
- Main points
- ChIP-exo (1)
- ChIP-exo (2)
- Reb1 (1)
- Reb1 (2)
- Phd1
- What ChIP-exo can do
- General transcription factors (GTFs)
- Application of ChIP-exo to transcription machinery
- TBP and TFIIB distribution around TATA boxes
- Other transcription factors around TATA boxes
- Most PICs contain all the GTFs
- Interpretation of TFIIB ChIP-exo patterns (1)
- Interpretation of TFIIB ChIP-exo patterns (2)
- Interpretation of TFIIB ChIP-exo patterns (3)
- The nature of "TATA-less" promoters
- PICs are mostly excluded from coding regions
- Intergenic regions have a multitude of PICs (1)
- Intergenic regions have a multitude of PICs (2)
- GTFs are rarely found in termination regions
- Conclusions
- Acknowledgements
Topics Covered
- Genomics of gene regulation
- Chromatin
- Canonical nucleosome organization around genes
- ChIP-exo mapping of sequence-specific transcription factors
- Genomic organization of transcription pre-initiation complexes
Links
Series:
Categories:
Talk Citation
Pugh, F. (2014, February 4). Genome-wide organization of chromatin and the transcription machinery [Video file]. In The Biomedical & Life Sciences Collection, Henry Stewart Talks. Retrieved December 3, 2024, from https://doi.org/10.69645/GIMU7149.Export Citation (RIS)
Publication History
Financial Disclosures
- Prof. Frank Pugh has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.
A selection of talks on Genetics & Epigenetics
Transcript
Please wait while the transcript is being prepared...
0:00
Hello.
Today's talk is on the
Genome-wide Organization
of Chromatin and the
Transcriptional Machinery.
My name is Dr. Frank Pugh.
I'm from the Center for Eukaryotic
Gene Regulation at Penn State
University in State
College, Pennsylvania.
As a way of background, I've
got my undergraduate degree
at Cornell University in
Ithaca, New York in 1983
and then went on to graduate
school in biochemistry
at the University of
Wisconsin at Madison,
working on genetic recombination.
I got my Ph.D. in 1987.
I then went on to the
University of California
at Berkeley where I studied under
Dr. Robert Teigen for a postdoc
After which in 1991, I started my
own lab at Penn State University
and have been there ever since.
And the focus of my research has
been on biochemical and genomic
mechanisms of eukaryotic
gene regulation.
0:56
Shown here is an image of
a typical eukaryotic gene
and the proteins that bind to it.
The green balls that you
see are the nucleosomes
which package the chromatin.
And you can see by that
small black arrow, the TSS,
is where the transcriptional
start site resides.
Between the minus 1 and
the plus 1 nucleosomes
is an open region where
there are no nucleosomes.
And that is where the
transcription machinery assembles.
And there is, perhaps, four stages
that you can think of assembly.
One is orchestration.
That's those red circles
that bind to specific DNA
sequences at or near
the minus 1 nucleosome.
The second step involves,
perhaps, chromatin remodeling,
the rearrangement of
proteins on the DNA surface
to make the DNA more accessible to
other transcription factor binding.
So that's step two, access.
The third step is the assembly
of the general transcriptional
machinery in the initiation phase.
That's shown in light
blue at the promoter
nucleosome-free region.
And then, that's
followed in step four
by the recruitment of RNA
polymerase in elongation factors
that ultimately need
to enter into the gene
and transcribe the genome.
We're going to first talk
about the organization
of nucleosomes shown here.
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