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Title [sv]
Undersökning av en ny funktion hos humant DNA polymerase theta (POLQ)
Title [en]
Investigation of a novel function of human DNA Polymerase theta (POLQ)
Abstract [sv]
DNA polymerase theta (POLQ) is one of 16 DNA polymerases in human cells. The function of POLQ is still unclear but it has been suggested that POLQ participates in DNA repair pathways. One interesting aspect of POLQ is its homology to high fidelity Family A DNA polymerases yet it replicates DNA with low fidelity and has trans-lesion synthesis properties similar to Family X polymerases. We have found that POLQ has yet another unique property, the capacity to carry out template independent DNA synthesis in a manner similar to that of terminal deoxyribonucleotide transferase (TdT). Both multiple and single turnover kinetics assays will be used to establish baseline values for the nucleotide preference, DNA length requirements and metal cofactor requirements of this novel function. We will also investigate the ability of POLQ to utilize substrates such as mismatched and frayed primer termini as these structures may result from DNA synthesis during damage repair and the activity of error prone DNA polymerases. We will use site-directed mutagenesis to try to identify mutants of POLQ that have lost the capacity to extend single-stranded DNA, but can still synthesize DNA with the help of a template strand. These mutants will be expressed in cell lines to explore if template independent DNA synthesis by POLQ has any biological relevance. If so, this property of POLQ might be an ideal drug target to screen for inhibitors that could increase the efficacy of current cancer treatments
Principal InvestigatorJohansson, Erik
Coordinating organisation
Umeå University
Funder
Period
2011-01-01 － 2013-12-31
Identifiers
DiVA, id: project:1081Project, id: 2010-03558_VR

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