Factores de replicación y combinación que contribuyen a la

reparación del DNA

Sergio UribePrograma de Doctorado en Ciencias Médicas

Universidad Austral de Chile

Reparación del DNA

Development of cancer stem cells from the normal stem cells and progenitor cells. Accumulation of DNA errors in normal stem cells or progenitor cells are activated to generate a cancer stem cells (CSCs) that further generate a primary tumor constituting CSCs and other tumor cells. Vaish Molecular Cancer 2007 6:26 doi:10.1186/1476-4598-6-26

From: DNA Repair

Copyright © 2002, Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, and Peter Walter; Copyright © 1983, 1989, 1994, Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, and James D. Watson .

From: DNA Repair

Copyright © 2002, Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, and Peter Walter; Copyright © 1983, 1989, 1994, Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, and James D. Watson .

From: DNA Repair

Copyright © 2002, Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, and Peter Walter; Copyright © 1983, 1989, 1994, Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, and James D. Watson .

From: DNA Repair

Copyright © 2002, Bruce Alberts, Alexander Johnson, Julian Lewis, Martin Raff, Keith Roberts, and Peter Walter; Copyright © 1983, 1989, 1994, Bruce Alberts, Dennis Bray, Julian Lewis, Martin Raff, Keith Roberts, and James D. Watson .

Materials and methods

Yeast strains and plasmids

Growing conditions, cell cycle arrests, and drug treatments

Spot assays of drug sensitivity

Protein techniques

FACS analysis

Extraction of replication intermediates and the 2D gel procedure

Quantification of replication intermediates

Materials and methods

Results

Results The role of the HR factors Rad55 and Rad59 in the

formation of template switch intermediates Rad55, but not Rad55 phosphorylation by Rad53 or

Rad59, is required for template switch replication The effect of the RPA mutation, rfa1-t11, on template

switch RPA, promoting the strand invasion step of homologous

recombination, is required for template switch replication

The Exo1 exonuclease is required for efficient damage-induced template switch events

Exo1 contributes to damage-bypass replication by template switch

TLS polymerases are not required for template switch–mediated damage bypass Translesion synthesis polymerases do not

contribute to the DNA synthesis step of template switch.

Differential requirements for replicative polymerases in template switch replication Polδ but not Polε, is required for template switch

replication

...understanding how different DNA synthetic and repair demands are orchestrated to prevent the accumulation of DNA damage and maintain chromosomal stability has important implications for enhancing our knowledge of how cells are protected from cancer-causing alterations

Referencias

Maresca M, Erler A, Fu J, Friedrich A, Zhang Y, Stewart AF. Single-stranded heteroduplex intermediates in lambda Red homologous recombination. BMC Molecular Biology 2010, 11:54

Vanoli F, Fumasoni M, Szakal B, Maloisel L, Branzei D. Replication and recombination factors contributing to recombination-dependent bypass of DNA lesions by template switch. PLoS Genet. 2010 Nov 11;6(11):e1001205.

Alberts B et al. Molecular Biology of the Cell, 5th edition. Ch 5. DNA Replication, Repair, and Recombination

Vaish M. Mismatch repair deficiencies transforming stem cells into cancer stem cells and therapeutic implications. Mol Cancer. 2007 Apr 2;6:26.