New insights into the GINS complex explain the controversy between existing structural models

Marta Carroni; Matteo De March; Barbara Medagli; Ivet Krastanova; Ian A. Taylor; Heinz Amenitsch; Hiroyuchi Araki; Francesca M. Pisani; Ardan Patwardhan; Silvia Onesti

doi:10.1038/srep40188

New insights into the GINS complex explain the controversy between existing structural models

Marta Carroni, Matteo De March, Barbara Medagli, Ivet Krastanova, Ian A. Taylor, Heinz Amenitsch, Hiroyuchi Araki, Francesca M. Pisani, Ardan Patwardhan, Silvia Onesti^*

^*Corresponding author for this work

Institute of Inorganic Chemistry (6330)

Research output: Contribution to journal › Article › peer-review

Abstract

GINS is a key component of eukaryotic replicative forks and is composed of four subunits (Sld5, Psf1, Psf2, Psf3). To explain the discrepancy between structural data from crystallography and electron microscopy (EM), we show that GINS is a compact tetramer in solution as observed in crystal structures, but also forms a double-tetrameric population, detectable by EM. This may represent an intermediate step towards the assembly of two replicative helicase complexes at origins, moving in opposite directions within the replication bubble. Reconstruction of the double-tetrameric form, combined with small-angle X-ray scattering data, allows the localisation of the B domain of the Psf1 subunit in the free GINS complex, which was not visible in previous studies and is essential for the formation of a functional replication fork.

Original language	English
Article number	40188
Journal	Scientific Reports
Volume	7
DOIs	https://doi.org/10.1038/srep40188
Publication status	Published - 10 Jan 2017

ASJC Scopus subject areas

General
Materials Science(all)

Fields of Expertise

Human- & Biotechnology

Treatment code (Nähere Zuordnung)

Basic - Fundamental (Grundlagenforschung)
Experimental

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title = "New insights into the GINS complex explain the controversy between existing structural models",

abstract = "GINS is a key component of eukaryotic replicative forks and is composed of four subunits (Sld5, Psf1, Psf2, Psf3). To explain the discrepancy between structural data from crystallography and electron microscopy (EM), we show that GINS is a compact tetramer in solution as observed in crystal structures, but also forms a double-tetrameric population, detectable by EM. This may represent an intermediate step towards the assembly of two replicative helicase complexes at origins, moving in opposite directions within the replication bubble. Reconstruction of the double-tetrameric form, combined with small-angle X-ray scattering data, allows the localisation of the B domain of the Psf1 subunit in the free GINS complex, which was not visible in previous studies and is essential for the formation of a functional replication fork.",

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AU - Carroni, Marta

AU - De March, Matteo

AU - Medagli, Barbara

AU - Krastanova, Ivet

AU - Taylor, Ian A.

AU - Amenitsch, Heinz

AU - Araki, Hiroyuchi

AU - Pisani, Francesca M.

AU - Patwardhan, Ardan

AU - Onesti, Silvia

PY - 2017/1/10

Y1 - 2017/1/10

N2 - GINS is a key component of eukaryotic replicative forks and is composed of four subunits (Sld5, Psf1, Psf2, Psf3). To explain the discrepancy between structural data from crystallography and electron microscopy (EM), we show that GINS is a compact tetramer in solution as observed in crystal structures, but also forms a double-tetrameric population, detectable by EM. This may represent an intermediate step towards the assembly of two replicative helicase complexes at origins, moving in opposite directions within the replication bubble. Reconstruction of the double-tetrameric form, combined with small-angle X-ray scattering data, allows the localisation of the B domain of the Psf1 subunit in the free GINS complex, which was not visible in previous studies and is essential for the formation of a functional replication fork.

AB - GINS is a key component of eukaryotic replicative forks and is composed of four subunits (Sld5, Psf1, Psf2, Psf3). To explain the discrepancy between structural data from crystallography and electron microscopy (EM), we show that GINS is a compact tetramer in solution as observed in crystal structures, but also forms a double-tetrameric population, detectable by EM. This may represent an intermediate step towards the assembly of two replicative helicase complexes at origins, moving in opposite directions within the replication bubble. Reconstruction of the double-tetrameric form, combined with small-angle X-ray scattering data, allows the localisation of the B domain of the Psf1 subunit in the free GINS complex, which was not visible in previous studies and is essential for the formation of a functional replication fork.

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New insights into the GINS complex explain the controversy between existing structural models

Abstract

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