# Structural studies of function and regulation of microtubules and transcriptional gene expression machinery

> **NIH NIH R35** · UNIVERSITY OF CALIFORNIA BERKELEY · 2024 · $481,500

## Abstract

Project Abstract
 Our lab is dedicated to the mechanistic understanding of macromolecular function through the visualization of
structure, dynamics, and regulatory interactions. Towards that goal, we use cryo-EM, together with biochemical
and biophysical assays. Our areas of study are centered on the characterization of the regulatory molecular
mechanisms governing the function of microtubules and of human transcription/epigenetic complexes.
 Microtubules (MTs) are essential polymers in eukaryotic cells s built of -tubulin dimers. Dynamic instability,
the switching between growing and shrinking phases due to the coupling of the assembly process to the exchange
and hydrolysis of GTP in -tubulin, is an essential property for MT function. Many MT cellular partners modulate
MT dynamics or utilize it to carry out specific functions. In the past. In the last 5 years, we confirmed and
complemented our past studies, which used non-hydrolyzable GTP analogs to characterize conformational
changes in MTs that accompany GTP hydrolysis, now using of GTP-hydrolysis tubulin mutants. We also defined
the mode of binding and action of cellular factors that regulate MT assembly, dynamics and organization in the
cell and described the effect of tubulin acetylation on MT structure and function. We will continue this work with
the central theme of adding complexity to our studies in order to bring us closer to the regulated function of
microtubule cellular systems as we also add techniques complementary to our major tool, cryo-EM.
 Transcriptional regulation of gene expression is critical for growth and survival, and of obvious significance to
human health. Its initiation involves RNA polymerase II (Pol II) together with TFIIA, TFIIB, TFIID, TFIIE, TFIIF, and
TFIIH. Regulation is achieved by sequence-specific activators or repressors, co-factors, and chromatin
remodeling/modifying complexes. During the last funding period we defined the structures of human TFIID[17] and
TFIIH[18], as well as other large transcriptional coactivators like the human SAGA[19], and the yeast NuA4 (in
preparation) and RSC complexes[20]. We will now characterize the human TIP60 complex, the binding of
transcriptional co-activators to chromatin substrates, and pursue the dynamic visualization of TFIID engagement
with promoter DNA to gain further knowledge of how these complexes work in the nucleus to regulate gene
expression.
 Polycomb repressive complex 2 (PRC2) is an epigenetic gene silencer that methylates lysine 27 of histone H3
and is essential for cellular differentiation and development. After obtaining the structure of human PRC2 with its
cofactors JARID2 and AEBP2 and its interaction with a di-nucleosome, we have now defined how PRC2
recognizes mono-ubiquitylated nucleosome, the substrate created by the other major Polycomb complex, PRC1.
We will now further characterize the regulatory landscape of PRC2 looking into other histone modifications,
different forms of PRC2 (i.e., other ...

## Key facts

- **NIH application ID:** 10904741
- **Project number:** 5R35GM127018-07
- **Recipient organization:** UNIVERSITY OF CALIFORNIA BERKELEY
- **Principal Investigator:** Eva Nogales
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $481,500
- **Award type:** 5
- **Project period:** 2018-05-01 → 2028-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10904741

## Citation

> US National Institutes of Health, RePORTER application 10904741, Structural studies of function and regulation of microtubules and transcriptional gene expression machinery (5R35GM127018-07). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10904741. Licensed CC0.

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