| |
 |
UNIVERSITY OF BUCHAREST
FACULTY OF PHYSICS Guest
2024-11-23 17:59 |
 |
|
|
|
Conference: Bucharest University Faculty of Physics 2017 Meeting
Section: Biophysics; Medical Physics
Title:
Convenient method for measuring transition times between interchanging protein DNA complex by tethered particle motion technique
Authors:
Elena IONITA(1,2), Octavian G. DULIU(2)
*
Affiliation:
1) Horia Hulubei National Institute for Physics and Nuclear Engineering (IFIN-HH), Department of Life and Environmental Physics, Bucharest-Magurele 077125, Romania
2) University of Bucharest, Faculty of Physics, PO Box MG-38, Bucharest-Magurele, Romania
E-mail
elena_ionita44@yahoo.com
Keywords:
V(D)J Recombination, RAG-recombination activing gene protein, RSS recognition signal sequence,TPM.
Abstract:
In all vertebrates, including humans, the immune system uses V(D)J recombination to generate the diverse repertoire of B and T-cells antigen receptors. The genes encoding the variable region of the receptor are assembled from small segments (V, D and J) by two lymphoid specific RAG 1 and 2 proteins (recombination activating gene products). For initiate the process of recombination, RAG binds two specific DNA sites- recombination signal sequences (RSS). RSSs consists by an palindromic heptamer and a nonamer elements separated by a non-conserved spacer of either 12 or 23 bp which gives their name of 12 or 23RSS. Many enzymatic and mechanistic issues are still poorly understood in this field. A very important unanswered question in our field addresses the nature of the process and rates of formation or dissociation of RAG-RSS binding reaction of 12SC, 23SC or of the PC formation. Using the TPM we detected transition between different state of DNA-RAG interaction. These experiments add a new profound dimension to the study of the protein-DNA dynamics occurring during V(D)J recombination.
References:
1. Daniel J. Schlingmana, Andrew H. Mackb, Simon G.J. Mochrieb, and Lynne Regana, A new method for the covalent attachment of DNA to a surface
for single-molecule studies, Colloids Surf. B Biointerfaces. 2011 March ; 83(1): 91–95. doi:10.1016/j.colsurfb.2010.11.002.
2. S.G.J. Mochrie, A.H. Mack, et al., L. Regan unwinding and rewinding the nucleosome inner turn: force dependence of the kinetic rate constants Phys. Rev. E Stat. Nonlin. Soft Matter Phys., 87 (2013), p. 012710
3. A.H. Mack, D.J. Schlingman, et al., S.G.J. Mochrie practical axial optical trapping Rev. Sci. Instrum., 83 (2012) 103106–103106
Acknowledgement:
We thank Peter Koo member of Simon Mochrie Lab, Yale University, Departament of Physics, for matlab development:
http://adsabs.harvard.edu/abs/2015PhDT.......301K
ProQuest Dissertations And Theses; Thesis (Ph.D.)--Yale University, 2015.; Publication Number: AAT 10009717; ISBN: 9781339461588; Source: Dissertation Abstracts International, Volume: 77-06(E), Section: B.; 219 p.
|
|
|
|