Structural study of the assembly of human TRF2/RAP1 telomeric complex

PhD in Biochemistry and Structural Biology

  • Prepared under the supervision of Dr Marie-Hélène Le Du in the team “Nuclear envelope, telomeres and DNA repair” of the Department of Biochemistry, Biophysics and Structural Biology (B3S) of the Institute of Integrative Biology of the Cell (I2BC), UMR 9198 CNRS, CEA, Université Paris-Sud.
  • Publicly defended on September 22nd 2015 at the Institut National des Sciences et Techniques du Nucléaire (INSTN), Saclay, France.
  • Thesis prize awarded by the French Crystallographic Association in 2016.

Abstract

Telomeres are the ends of eukaryotic linear chromosomes. They are made of tandem repeats of a short guanine-rich motif and bound by specific proteins. In vertebrates, these proteins form a complex called shelterin, the integrity of which is critical to ensure proper replication of chromosome ends and to protect them against illicit targeting by DNA double-strand break repair pathways. Telomere dysfunctions lead to genome instability, which can ultimately cause senescence or cancer. Telomeres are a subnuclear region in which shelterin proteins are highly enriched, enhancing low affinity interactions of important biological function. Among shelterin proteins, telomeric repeat-binding protein TRF2 and its constitutive partner RAP1 are the main factors responsible for end protection. We studied in details the assembly of TRF2/RAP1 complex by means of integrated structural, biophysical and biochemical approaches. We showed that this assembly displays important conformational adjustments of both proteins, and involves a low affinity interaction engaging large regions in both proteins which affects their interaction properties.

Keywords: telomeres, TRF2, RAP1, SAXS, ITC, crystallography, protein footprinting.

Full text

The thesis was defended in a French university and is therefore written in French. The final version of the manuscript is available through the Thèses en ligne public archive.

Here is a self-archived version which has the final published version of the article instead of a draft (only difference with the officially deposited version above), and here is its OpenPGP digital signature. To verify the signature with GnuPG, run the following command in the directory that contains both the PDF file and the signature file:

gpg --verify these-Guillaume-Gaullier-depot-final.pdf.asc

You should be able to find the following information in the output (otherwise this means either the PDF or signature file got altered in some way):

  • signed on Friday March 4th 2016
  • with RSA key 0x4599AF1249AC270E
  • good signature (look for this information first, the rest is only here to confirm the signature)
  • primary key fingerprint: F918 EF6C 9061 BAD6 6C84 0A8A B111 2808 C99D 8831
  • subkey fingerprint: CC5E B20F B0AC FAE5 41CB 70EB 4599 AF12 49AC 270E

All source files and full modification history are available as a public git repository.

Some day, I may also put online a PDF file of the slides used for the defense.