Germline minisatellite instability in the presence and absence of MLH1, a mismatch repair protein

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en
dc.contributor.advisor Kauppi, Liisa Shrestha, Kul 2014-10-03T07:22:26Z 2014-10-03T07:22:26Z 2014-09-29
dc.description.abstract MLH1 (MutL homolog 1) is a mismatch repair protein that can form heterodimer with MLH3 in meiosis. A functional MLH1-MLH3 heterodimer is essential for crossover recombination. Not much is known about the effect of loss of MLH1 function on mini satellite instability. The aim of this project is to investigate mini satellite instability in germ line cells of wild-type and Mlh1-/- male mice of the C57BL/6 strain, with a particular focus on mini satellite locus Prdm9 (a meiotic recombination protein in metazoans). An attempt is made to find mini satellites that function as potential DNA binding sites for PRDM9 and mini satellite instability analysis is also performed for one of them (Ms-X165.369). Prdm9 locus is compared to a known stable and an unstable mini satellite, Ms-X165.369 and two characterized microsatellites. Spermatocytes were isolated from mouse testis based on their size, and DNA was extracted. As a control, DNA extracted from somatic tissue was used. Small-pool PCR followed by Southern blotting was used to investigate and compare stability of the different mini satellite loci. In silico analysis was conducted to predict DNA binding motifs and DNA binding affinities for newly identified Prdm9 variants. We found germ line instability at Prdm9 locus to be higher in Mlh1-/- mouse than in wild type mouse, with germ line instability of 16.6% and 1.4% per gamete respectively. No somatic mutation was observed at Prdm9 locus for both the genotype. At Ms-X165.369 locus, neither germ line nor somatic mutation was observed in either of the genotype. All the PRDM9 variants were predicted to have different DNA binding motifs. DNA binding affinity of PRDM9 variants to Ms-X165.369 was predicted to increase by at most 39% and decrease by at least 51.5% and to the progenitor PRDM9's DNA binding motif by at least 68%, compared to the progenitor PRDM9. Our results show that MLH1 is required for germ line mini satellite stability. en
dc.format.extent 69 + 8
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.title Germline minisatellite instability in the presence and absence of MLH1, a mismatch repair protein en
dc.type G2 Pro gradu, diplomityö en Perustieteiden korkeakoulu fi
dc.subject.keyword germline minisatellite instability en
dc.subject.keyword Prdm9 en
dc.subject.keyword Ms-X165.369 en
dc.subject.keyword mismatch repair en
dc.subject.keyword MLH1 en
dc.subject.keyword single-molecule PCR en
dc.identifier.urn URN:NBN:fi:aalto-201410042717
dc.programme.major Computational Systems Biology fi
dc.programme.mcode IL3013 fi
dc.type.ontasot Master's thesis en
dc.type.ontasot Diplomityö fi
dc.contributor.supervisor Rousu, Juho
dc.programme Master's Degree Programme in Computational and Systems Biology (euSYSBIO) fi

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