Cell Biology
Catherine M. Green
Cancer Research UK Career Development Fellow
Email: cmg1003@cam.ac.uk
Tel.: +44 (0)1223 769353
Replication Fork Dynamics - Protein-protein interactions during DNA replication
The replication of the genome must be performed completely and accurately once in each cell cycle in order to maintain genome stability and to minimise mutations that can lead to cancer development. The process of DNA replication comprises a series of enzymatic steps that must be temporally and spatially controlled within the cell nucleus if errors are to be avoided. We are investigating this control at a molecular level by analyzing interactions and associations between proteins that perform replicative functions both in vitro and in vivo.
We are a newly established lab, funded by Cancer Research UK, Our main focus is an investigation of protein traffic on a key replication protein, proliferating cell nuclear antigen (PCNA). PCNA is a ring shaped protein that encircles the template DNA and can slide along the linear DNA molecule. Any proteins that interact with PCNA are therefore brought in close proximity to the DNA. PCNA is known to interact with proteins essential for all stages of the replication process, including polymerases, nucleases, ligases and helicases. PCNA is also important for ensuring that chromatin and epigenetic marks are correctly assembled onto replicated DNA as well as for DNA repair events and also for controlling the time in the cell cycle at which replication is permitted. In fact more than 70 different proteins have been shown to interact with PCNA. Our overall aim is to understand how these multiple interactions can be constrained so that only the appropriate ones occur at any particular time and place.
To achieve this aim we use a variety of methods to analyse interactions between proteins. These include: In vitro analysis of interactions between replication proteins by surface plasmon resonance (SPR); in vitro and in vivo analyses of protein associations by fluorescence resonance energy transfer (FRET); and biochemical purification of replication protein complexes.
Selected publications
- Lehmann AR, Niimi A, Ogi T, Brown S, Sabbioneda S, Wing JF, Kannouche PL, Green CM, 2007. Translesion synthesis: Y-family polymerases and the polymerase switch. DNA Repair (Amst). Jul 1;6(7):891-9.
- Green CM., 2006. One ring to rule them all? Another cellular responsibility for PCNA. Trends Mol Med. Oct;12(10):455-8.
- Bienko M, Green CM, Crosetto N, Rudolf F, Zapart G, Coull B, Kannouche P, Wider G, Peter M, Lehmann AR, Hofmann K, Dikic I., 2005. Ubiquitin-binding domains in Y-family polymerases regulate translesion synthesis. Science. Dec 16;310(5755):1821-4.
- Albertella MR, Green CM, Lehmann AR, O'Connor MJ., 2005. A role for polymerase eta in the cellular tolerance to cisplatin-induced damage. Cancer Res. Nov 1;65(21):9799-806.
- Green CM, Lehmann AR, 2005. Translesion synthesis and error-prone polymerases. in Genome Instability in Cancer Development, E. A. Nigg, ed. ( Netherlands, Springer), pp. 199-223.
- Green CM, Almouzni G., 2003 Local action of the chromatin assembly factor CAF-1 at sites of nucleotide excision repair in vivo. EMBO Journal. Oct-01;22(19):5163-74.
- Gilbert CS, Green CM, Lowndes NF., 2001. Budding yeast Rad9 is an ATP-dependent Rad53 activating machine. Molecular Cell. Jul;8(1):129-36.
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