Search bioRxiv⌕ Search

Biology subjects

Dalgleish, J. L. T.

Publications and source records attributed to Dalgleish, J. L. T..

2 recordsLinked to original sources

FACT safeguards promoter topology by maintaining nucleosomes and restricting chromatin factor spreading

Facilitates chromatin transcription (FACT) is a histone chaperone that displaces and re-assembles histones during transcription. Recent studies have reported a minor role for FACT in chromatin architecture. We have recently shown that active gene promoters form nanoscale domains and proposed they are created by the biophysical properties of nucleosome-free regions. Here we use base-pair resolution Micro Capture-C ultra to show that, following FACT degradation, nanoscale domains are lost and subnucleosomal chromatin interactions are rearranged at active promoters. Nucleosome-free regions at these promoters expand and chromatin-binding factors invade the newly accessible chromatin, indicating FACT maintains the integrity of active promoters by opposing DNA-binding factor spreading into gene bodies. Finally, we show increased interactions between promoters across topologically associating domains, suggesting large-scale structural changes upon FACT loss. Thus, we demonstrate FACT plays a major role in chromatin organisation and provide in vivo evidence that nucleosomes drive both local and long-range chromatin architecture.

molecular biology↗

Cancer-associated DAXX mutations reveal a critical role for ATRX localization in ALT suppression

To maintain genome stability, proliferating cells must enact a program of telomere maintenance. While most tumors maintain telomeres through the action of telomerase, a subset of tumors utilize a DNA-templated process termed Alternative Lengthening of Telomeres or ALT. ALT is associated with mutations in the ATRX/DAXX/H3.3 histone chaperone complex, which is responsible for deposition of non-replicative histone variant H3.3 at heterochromatic regions of the genome including telomeres. We wished to better understand the role DAXX plays in ALT suppression, and to determine which disease-associated DAXX mutations are unable to suppress ALT. To answer this question, we have leveraged the G292 cell line, in which ATRX is wild type but DAXX has undergone a fusion event with the non-canonical kinesin KIFC3. Restoration of wild-type DAXX in G292 localizes ATRX and abrogates ALT. Using this model system, we tested the ability of a panel of disease-associated DAXX missense variants to suppress ALT. Missense mutations in the ATRX binding domain, the histone binding domain, and the C-terminal SUMO interaction motif reduce the ability of DAXX to suppress ALT. Unexpectedly, we find that mutations in the DAXX histone binding domain lead to failure of ATRX localization. We conclude that a key function of DAXX in ALT suppression is the localization of ATRX to nuclear foci.

cancer biology↗