Structural Characterization of the Saccharomyces cerevisiae THO Complex by Small-Angle X-Ray Scattering

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DOI

  • Jesper Buchhave Poulsen, Centre for mRNP Biogenesis and Metabolism, Aarhus University, Aarhus, Denmark., Danmark
  • Lee Edward Sanderson, Centre for mRNP Biogenesis and Metabolism, Aarhus University, Aarhus, Denmark., Danmark
  • Emil Dandanell Agerschou, Centre for mRNP Biogenesis and Metabolism, Aarhus University, Aarhus, Denmark., Danmark
  • Emil Dedic, Centre for mRNP Biogenesis and Metabolism, Aarhus University, Aarhus, Denmark., Danmark
  • Thomas Boesen
  • Ditlev E. Brodersen

The THO complex participates during eukaryotic mRNA biogenesis in coupling transcription to formation and nuclear export of translation-competent messenger ribonucleoprotein particles. In Saccharomyces cerevisiae, THO has been defined as a heteropentamer composed of the Tho2p, Hpr1p, Tex1p, Mft1p, and Thp2p subunits and the overall three-dimensional shape of the complex has been established by negative stain electron microscopy. Here, we use small-angle X-ray scattering measured for isolated THO components (Mft1p and Thp2p) as well as THO subcomplexes (Mft1p-Thp2p and Mft1p-Thp2p-Tho2p) to construct structural building blocks that allow positioning of each subunit within the complex. To accomplish this, the individual envelopes determined for Mft1p and Thp2p are first fitted inside those of the Mft1p-Thp2p and Mft1p-Thp2p-Tho2p complexes. Next, the ternary complex structure is placed in the context of the five-component electron microscopy structure. Our model reveals not only the position of each protein in the THO complex relative to each other, but also shows that the pentamer is likely somewhat larger than what was observed by electron microscopy.

OriginalsprogEngelsk
Artikelnummere103470
TidsskriftPLOS ONE
Vol/bind9
Nummer7
Sider (fra-til)1-11
Antal sider11
ISSN1932-6203
DOI
StatusUdgivet - jul. 2014

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