Ccctc-binding factor (ctcf): choreographer of genome organization

Project Grant Awarded Last action

Award ID
F32GM149128
Recipient
Awarding agency
Obligated
$69,080

Award facts

Assistance listing
93.859 Biomedical Research and Research Training
Completion date
Not on record
Place of performance
Netherlands

Award details come from USAspending.gov.

Description

As the agency reported it.

Abstract
Packaging 2M of human genomic DNA into the 10ΜM nucleus requires formation of a three-dimensional structure that is highly compact, yet flexible. The spatial and temporal structure of chromatin regulates gene expression, development, and DNA maintenance. Disruption of genomic architecture is associated with developmental disorders and cancer. At the mesoscopic level, chromosomes are divided into topologically associated domains (TAD), the borders of which ccctc-binding factor (ctcf) is found to be enriched. Described as 'master weaver of the genome', ctcf is a multivalent protein of 11 zinc fingers that drive directional binding to ~60,000 ctcf-binding sites (CBS) across the human genome. Recent findings suggest a model where the cohesin motor protein extends loops until encountering convergent ctcf- bound cbss which stabilize the established loop. While the model is promising, this mechanism has not yet been observed in action. The question of how genomic structures are efficiently remodelled in response to regulatory signals remains unaddressed by the loop extrusion model. Dynamic reorganization of genomic structure includes the dramatic condensation of chromosomes that occurs with the cell cycle as well reorganization that is necessary to provide access to the machinery of gene expression, DNA replication and repair. As a driver of chromosomal structure, ctcf:DNA complexes must be similarly dynamic to allow the necessary structural flexibility. The main goal of the proposed research is to determine how chromosomal structures established or maintained by ctcf are stable, but also responsive to signals triggering rearrangement. To this end, the hypothesis that binding of ctcf to CBS-containing DNA results in the adoption of a conformationally dynamic ctcf:DNA complex, the dynamics of which are the target of regulation, will be tested. The formation and conformational dynamics of the ctcf:DNA complex will be characterized in aim 1 using optical tweezers with fluorescence microscopy and atomic force microscopy (AFM). The effects of ctcf activity modulators on complex formation and dynamics will be elucidated in aim 2 by addressing the impact of alternating the CBS consensus sequence and phosphomimic mutation of ctcf. Ctcf association with mitotic chromosomes will be mapped correlated to its effect on chromosome architecture in aim 3, using optical tweezers to trap unfixed mitotic chromosomes, imaging associated fluorescent ctcf, and interrogating with force. Completion of the proposed work will yield an understanding of ctcf:DNA complex dynamics and their role in maintaining dynamic genomic structures and training in cutting-edge techniques combining super-resolution fluorescence microscopy and the application of force. Supervised by DR. Gijs wuite in the physics of living systems section at vrije universiteit amsterdam, training will prepare the applicant for a to found their own research group investigating dynamic processes in the regulation of DNA maintenance.

Transactions

Action Obligated Date
Amendment -$12,200
Amendment -$56,880
Amendment $12,200
Amendment $12,200
Amendment $56,880
Original award $56,880

Quick answers

Who received award F32GM149128?

F32GM149128 was awarded to Stichting VU by National Institute of General Medical Sciences (National Institutes of Health) on March 22, 2024.

How much is award F32GM149128 worth?

$69,080 has been obligated on F32GM149128.

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