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HJMIGRA

Single-molecule analysis of Holliday-junction (HJ) migration by the human double-HJ dissolvasome

Total Cost €

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EC-Contrib. €

0

Partnership

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 HJMIGRA project word cloud

Explore the words cloud of the HJMIGRA project. It provides you a very rough idea of what is the project "HJMIGRA" about.

mechanism    bloom    total    investigates    supports    microscopy    subunits    holliday    molecule    labeling    roles    dissolvasome    structure    helicases    rearrangements    free    consists    combined    homologous    biochemical    human    crossover    action    position    dissolves    questions    regulatory    complex    breaks    recombination    resolved    group    processive    microfluidics    underlying    proteins    convergent    follow    chromosomal    hj    processed    family    error    dna    techniques    generate    humans    leads    understand    top3a    migration    rmi1    substrate    previously    btr    template    reflection    fluorescently    tirf    biophysical    stranded    independent    single    mechanisms    genome    ends    syndrome    double    provides    inaccessible    migrate    blm    mobile    fluorescence    rmi2    junction    exact    hjs    enzymatic    maintenance    molecular    engagement    solution    bacteriophage    dissolved    branch    solely    broken    final    critical    elucidation    decatenation    specialized    lambda    internal    helicase    components    outlined    hr    topoisomerases    recq    concerted    dsbs    dhj    dissolution    cleavage    hemicatenate    repair   

Project "HJMIGRA" data sheet

The following table provides information about the project.

Coordinator
EOTVOS LORAND TUDOMANYEGYETEM 

Organization address
address: EGYETEM TER 1-3
city: BUDAPEST
postcode: 1053
website: www.elte.hu

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Hungary [HU]
 Project website http://mk-lab.org
 Total cost 146˙239 €
 EC max contribution 146˙239 € (100%)
 Programme 1. H2020-EU.1.3.2. (Nurturing excellence by means of cross-border and cross-sector mobility)
 Code Call H2020-MSCA-IF-2014
 Funding Scheme MSCA-IF-EF-RI
 Starting year 2015
 Duration (year-month-day) from 2015-05-01   to  2017-04-30

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    EOTVOS LORAND TUDOMANYEGYETEM HU (BUDAPEST) coordinator 146˙239.00

Map

 Project objective

The project outlined here investigates the molecular mechanisms of the critical final steps of homologous recombination (HR) based DNA repair, a pathway that supports the error-free repair of double-stranded DNA breaks (DSBs). In HR, the broken DNA ends are processed and homologous DNA provides a template for repair. Engagement of both processed ends leads to the formation of a double Holliday-junction (DHJ) structure. DHJ can be resolved by enzymatic cleavage or dissolved by the concerted action of a specialized group of helicases (RecQ-family helicases including Bloom’s syndrome helicase (BLM)) and Type I topoisomerases (e.g. TOP3A). In humans the ‘dissolvasome complex’ consists of BLM, TOP3A and regulatory proteins (RMI1, RMI2), called the BTR complex. The BTR complex dissolves DHJ by 1. convergent branch migration of the two independent HJs and 2. decatenation of the final hemicatenate structure. Thus, dissolution solely results non-crossover products, which is necessary to avoid chromosomal rearrangements. What is the mechanism of HJ migration? What are the exact roles of the subunits of the BTR complex? How long can a HJ migrate (i.e. how processive is the ‘dissolvasome’)? How specific is the DHJ migration to the BTR complex compared to other human RecQ helicases? Here we aim to address these questions by using state-of-the-art single-molecule and solution biophysical and biochemical techniques. We will generate a previously inaccessible mobile HJ substrate integrated into λ-bacteriophage DNA. We will follow the processes underlying HJ migration by fluorescently labeling the BTR complex, HJ position and DNA end in total internal reflection fluorescence (TIRF) microscopy combined with microfluidics. Elucidation of the detailed roles of the BTR components in HJ branch migration will help us to understand their roles in genome maintenance.

 Publications

year authors and title journal last update
List of publications.
2016 Máté Gyimesi, Gábor M. Harami, Zsuzsa S. Kocsis, Mihály Kovács
Recent adaptations of fluorescence techniques for the determination of mechanistic parameters of helicases and translocases
published pages: 24-39, ISSN: 1046-2023, DOI: 10.1016/j.ymeth.2016.04.028
Methods 108 2019-07-23
2017 Gábor M. Harami, Yeonee Seol, Junghoon In, Veronika Ferencziová, Máté Martina, Máté Gyimesi, Kata Sarlós, Zoltán J. Kovács, Nikolett T. Nagy, Yuze Sun, Tibor Vellai, Keir C. Neuman, Mihály Kovács
Shuttling along DNA and directed processing of D-loops by RecQ helicase support quality control of homologous recombination
published pages: E466-E475, ISSN: 0027-8424, DOI: 10.1073/pnas.1615439114
Proceedings of the National Academy of Sciences 114/4 2019-07-23
2017 Mate Gyimesi, Zoltan Kovacs, Mihaly Kovacs
Holliday Junction Structure Development for Single-Molecule Visualization
published pages: 371a-372a, ISSN: 0006-3495, DOI: 10.1016/j.bpj.2016.11.2017
Biophysical Journal 112/3 2019-07-23

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