TY - JOUR
T1 - Combining electron microscopy with single molecule DNA fiber approaches to study DNA replication dynamics
AU - Vindigni, Alessandro
AU - Lopes, Massimo
N1 - Funding Information:
We would like to thank Ralph Zellweger for its contribution in the preparation of Fig. 3. The work in the A.V. laboratory is supported by NIH grant R01GM108648 and by DOD BRCP Breakthrough Award BC151728. The work in the M.L. laboratory is supported by the SNF grants 31003A_169959 and 31003A_146924, and by the ERC Consolidator Grant 617102.
Publisher Copyright:
© 2016 Elsevier B.V.
PY - 2017/6
Y1 - 2017/6
N2 - Replication stress is a crucial driver of genomic instability. Understanding the mechanisms of replication stress response is instrumental to improve diagnosis and treatment of human disease. Electron microscopy (EM) is currently the technique of choice to directly visualize a high number of replication intermediates and to monitor their remodeling upon stress. At the same time, DNA fiber analysis is useful to gain mechanistic insight on how genotoxic agents perturb replication fork dynamics genome-wide at single-molecule resolution. Combining these techniques has proven invaluable to achieve a comprehensive view of the mechanisms that ensure error-free processing of damaged replication forks. Here, we review how EM and single-molecule DNA fiber approaches can be used together to shed light into the mechanisms of replication stress response and discuss important cautions to be taken into account when comparing results obtained by EM and DNA fiber.
AB - Replication stress is a crucial driver of genomic instability. Understanding the mechanisms of replication stress response is instrumental to improve diagnosis and treatment of human disease. Electron microscopy (EM) is currently the technique of choice to directly visualize a high number of replication intermediates and to monitor their remodeling upon stress. At the same time, DNA fiber analysis is useful to gain mechanistic insight on how genotoxic agents perturb replication fork dynamics genome-wide at single-molecule resolution. Combining these techniques has proven invaluable to achieve a comprehensive view of the mechanisms that ensure error-free processing of damaged replication forks. Here, we review how EM and single-molecule DNA fiber approaches can be used together to shed light into the mechanisms of replication stress response and discuss important cautions to be taken into account when comparing results obtained by EM and DNA fiber.
UR - http://www.scopus.com/inward/record.url?scp=85007524957&partnerID=8YFLogxK
U2 - 10.1016/j.bpc.2016.11.014
DO - 10.1016/j.bpc.2016.11.014
M3 - Article
C2 - 27939387
AN - SCOPUS:85007524957
SN - 0301-4622
VL - 225
SP - 3
EP - 9
JO - Biophysical Chemistry
JF - Biophysical Chemistry
ER -