Functional analysis of Hsh155/SF3b1 interactions with the U2 snRNA/branch site duplex

RNA
Tucker J CarrocciAaron A Hoskins

Abstract

SF3b1 is an essential component of the U2 snRNP implicated in branch site (BS) recognition and found to be frequently mutated in several human cancers. While recent structures of yeast and human SF3b1 have revealed its molecular architecture, the importance of specific RNA:protein contacts and conformational changes remains largely uncharacterized. Here, we performed mutational analysis of yeast SF3b1, guided by recent structures of the spliceosome. We find that conserved amino acids contacting the U2 snRNA backbone of the U2/BS duplex are nonessential, and that yeast can tolerate truncation of the HEAT repeats containing these amino acids. The pocket housing the branchpoint adenosine (BP-A) is also amenable to mutation despite strong conservation. However, mutations that support viability can still lead to defects in splicing pre-mRNAs with nonconsensus BS substitutions found at -3, -2, -1, and +1 positions relative to the BP-A or at the branchpoint position. Through the generation of yeast and human chimeric proteins, we further defined the functionally conserved regions of Hsh155 as well as identify changes in BS usage resulting from inclusion of human SF3b1 HEAT repeats. Moreover, these chimeric proteins confer a sensitivit...Continue Reading

References

Jul 22, 1998·Molecular and Cellular Biology·O GozaniR Reed
Jul 10, 2002·The Journal of Biological Chemistry·An BoudrezMathieu Bollen
Nov 14, 2002·Annual Review of Genetics·David A Brow
Oct 27, 2007·Biochemical and Biophysical Research Communications·Chor-Wai LoMinoru Yoshida
Nov 21, 2007·RNA·Daniel J CrawfordMelissa J Moore
Feb 26, 2009·Cell·Markus C WahlReinhard Lührmann
Mar 12, 2011·Science·Aaron A HoskinsMelissa J Moore
Oct 11, 2011·The FEBS Journal·Akira YokoiYoshiharu Mizui
Apr 7, 2012·Trends in Biochemical Sciences·Aaron A Hoskins, Melissa J Moore
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Oct 17, 2012·Molecular and Cellular Biology·Hsueh-Lien Liu, Soo-Chen Cheng
Nov 6, 2012·Nature Reviews. Drug Discovery·Sophie BonnalJuan Valcárcel
Dec 5, 2013·The Journal of Biological Chemistry·Kerstin A EffenbergerMelissa S Jurica
Feb 18, 2014·Current Opinion in Structural Biology·Weijun Chen, Melissa J Moore
Jan 7, 2015·Genome Research·Tim R MercerJohn S Mattick
Jun 11, 2016·Nature Reviews. Cancer·Heidi DvingeRobert K Bradley
Jul 22, 2016·Wiley Interdisciplinary Reviews. RNA·Kerstin A EffenbergerMelissa S Jurica
Jul 23, 2016·Science·Chuangye YanYigong Shi
Aug 27, 2016·Science·Reinhard RauhutReinhard Lührmann
Apr 5, 2017·Trends in Genetics : TIG·Jermaine L Jenkins, Clara L Kielkopf
May 23, 2017·Nature·Clemens PlaschkaKiyoshi Nagai
Aug 10, 2017·Nature Reviews. Molecular Cell Biology·Lydia HerzelKarla M Neugebauer

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Citations

Nov 21, 2019·Critical Reviews in Biochemistry and Molecular Biology·Clarisse van der Feltz, Aaron A Hoskins
Mar 7, 2020·Cellular and Molecular Life Sciences : CMLS·Chengfu Sun
Feb 7, 2021·RNA·Clarisse van der FeltzAaron A Hoskins
Aug 6, 2021·Nature·Zhenwei ZhangReinhard Lührmann

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