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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.

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

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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