Isolation and Characterization of Separation of Function Mutations in the Saccharomyces Cerevisiae MLH1 Mismatch Repair Gene PDF Download
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Author: Sumeet Sarin
Publisher:
ISBN:
Category :
Languages : en
Pages : 64
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Author: Sumeet Sarin
Publisher:
ISBN:
Category :
Languages : en
Pages : 64
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Author: Marc Paul Waase
Publisher:
ISBN:
Category :
Languages : en
Pages : 74
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Author: Juan Argueso Almeida
Publisher:
ISBN:
Category :
Languages : en
Pages : 334
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Author: Barbara Studamire
Publisher:
ISBN:
Category :
Languages : en
Pages : 408
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Author: Julie Akiko Heck
Publisher:
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Category :
Languages : en
Pages : 228
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Author: Aaron J. Plys
Publisher:
ISBN:
Category :
Languages : en
Pages : 176
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Book Description
Replication errors that escape DNA polymerase proof-reading activity are efficientl y recognized and repaired by conserved DNA mismatch repair factors. The overall result is a drastic reduction in deletion mutations. The mechanistic details of how mismatch repair proteins execute mismatch removal have not been elucidated. The aim of my thesis is to better understand how mismatch repair factors interact with DNA in order to identify mismatch sites. My work reveals that the mismatch repair complex, MLH1-PMS1, has unique DNA diffusion characteristics facilitated by structural features of the two subunits. Through bulk assays and total internal reflectance fluorescence microscopy (TIRFM), I found that MLH1PMS1 could independently bind DNA and rapidly diffuse using the thermal energy of the system. Furthermore, MLH1-PMS1 was shown to be the first passively diffusing protein that could bypass stationary nucleosomes. In contrast, the DNA diffusion activity of the mismatch recognition complex MSH2-MSH6 was blocked by nucleosomes. The timing and nature of mismatch repair is linked with replication and is thus proposed that the differences seen for the two complexes have important implications for repair in the context of the chromatin state directly at the replication fork. Each subunit of the MLH1-PMS1 complex is composed of two defined globular domains connected by an unstructured linker arm. The linker arms of the complex are proposed to facilitate topological DNA binding and diffusion along DNA in a hopping/stepping mechanism. I found that TEV protease cleavage within the linker arms of MLH1-PMS1 disrupted DNA binding and mismatch repair in vitro and in vivo. Using a genetic mismatch repair assay I found that shortening of the linker arms in MLH1 had a drastic effect on function whereas similar changes in PMS1 had little or no effect. Purified truncated complexes were able to interact with DNA and form ternary complexes with MSH2-MSH6 at a mismatch. Future studies should focus on the diffusion characteristic for these complexes. Together, my work has important implications for understanding how mismatch repair proteins can rapidly identify their targets in a chromatin landscape.
Author:
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ISBN:
Category : Medicine
Languages : en
Pages : 1448
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Author: Paula E. Cohen
Publisher: S. Karger AG (Switzerland)
ISBN:
Category : Medical
Languages : en
Pages : 130
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This volume offers an extensive array of reviews on the roles of DNA repair proteins in meiotic recombination. The use of various organisms serves to underscore the marked degree of conservation of these processes among sexually reproducing organisms, but also highlights more subtle species-specific adaptations to meiotic recombination. Contributed by leaders in the field, each chapter reflects the current state of their research and also provides a forum for new ideas and concepts in meiotic recombination. By focussing on different pathways involved in meiosis and recombination, this volume highlights the complexity of these processes and demonstrates how much has been learned in recent years as a result of the multiple approaches to studying these events in various model organisms. Geneticists, cytogeneticists, developmental biologists, and those involved in research on infertility and germ cell biology will find much to interest them in this outstanding collection of reviews.
Author: Sam Thiagalingam
Publisher: Cambridge University Press
ISBN: 0521493390
Category : Mathematics
Languages : en
Pages : 597
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An overview of the current systems biology-based knowledge and the experimental approaches for deciphering the biological basis of cancer.
Author:
Publisher:
ISBN:
Category : Genetics
Languages : en
Pages : 770
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