Capturing Molecular Movies of DNA Replication and Repair
THUWAL, Kingdom of Saudi Arabia, June 11, 2013 /PRNewswire/ –
King Abdullah University of Science and Technology (KAUST) Assistant Professor of
Chemistry and Biology and Principal Investigator of the Laboratory of DNA Replication and
Recombination, Dr. Samir M. Hamdan, and his team, have published a groundbreaking paper on
the mechanism of flap endonuclease 1 (FEN1), in Cell Reports journal. The other authors of
the paper, entitled “Sequential and Multistep Substrate Interrogation Provides the
Scaffold for Specificity in Human Flap Endonuclease 1,” are: Mohamed A. Sobhy, Luay I.
Joudeh, Xiaojuan Huang, and Masateru Takahashi.
5′ nucleases, the superfamily to which FEN1 belongs,represent structure-specific
nucleases essential for DNA replication, repair, and recombination. Deficiencies in their
genes have been linked to several types of cellular stress and genomic instability.Their
outstanding puzzle is that they are highly conserved proteins, yet they recognize a
diverse range of RNA and DNA structures and cleave them primarily at the same position
relative to a 5′ end of a junction.
Structural studies propose a solution for this geometrical puzzle by capturing a DNA
bending intermediary step that position the 5′ end in the enzyme active pocket and unify
the 5′ nucleases cleavage site. However, these structures remain a static image of the DNA
bending intermediary step based on which speculation was made to address the most
important questions regarding how this intermediary step is induced and how 5′ nucleases
utilize the same intermediary step to recognize diverse range of substrates.
The team from KAUST’s Division of Biological and Environmental Sciences and
Engineering employed a sophisticated single molecule imaging technique, Forster resonance
energy transfer (smFRET), to capture “molecular movies” detailing the structure, dynamics,
and reaction mechanisms occurring during this process. The KAUST scientists were able to
build a timeline of up to seven intermediary steps before FEN1 commits to catalysis. Such
information cannot be accessed through conventional approaches.
These findings will influence how researchers think about the mechanism of other
members of 5′ nucleases and provide a new concept as to how biological macromolecules can
diversify their substrate specificity while maintaining a high degree of structural
Dr. Hamdan explained that the surprising finding is that FEN1 utilizes a highly
complex mechanism that sequentially verifies all substrate features before inducing the
superfamily unifying DNA bending intermediary. This sequential and multistep substrate
recognition process provides a scaffold that allows different 5′ nucleases to recognize
different substrates and restrict the induction of DNA bending to the last common step.
“We hope that our findings will serve as a base to design inhibitors against FEN1, whose
expression is highly correlated with tumor aggressiveness,” said Prof. Hamdan.
- "Sequential and Multistep Substrate Interrogation Provides the Scaffold for Specificity in Human Flap Endonuclease 1" [http://www.cell.com/cell-reports/fulltext/S2211-1247%2813%2900218-0 ] Cell Reports online - "A Scientific Oasis in a Desert" [http://news.cell.com/cellreports/cell-reports/a-scientific-oasis-in-a-desert ] Cell Reporter blog post by Dr. Hamdan - KAUST website [http://www.kaust.edu.sa ]
King Abdullah University of Science and Technology (KAUST) is an international,
graduate research university dedicated to advancing science and technology through
interdisciplinary research, education, and innovation. Students, faculty, scientists, and
engineers conduct fundamental and goal-oriented research to address the world’s pressing
scientific and technological challenges related to water, food, energy, and the
Located on the shores of the Red Sea, KAUST is the first LEED certified project in
Saudi Arabia and the largest LEED Platinum project in the world. In 2011, KAUST was named
Lab of the Year by leading science and technology publication, R&D Magazine.The following
year, the University’s Research Park was named the 2012 Emerging Research and Science Park
by the Association of University Research Parks (AURP). http://www.kaust.edu.sa
Samia Falimban, Head of University Communications, firstname.lastname@example.org,
SOURCE King Abdullah University of Science and Technology