English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Journal Article

Multiple protein-protein interactions converging on the Prp38 protein during activation of the human spliceosome.

MPS-Authors
/persons/resource/persons16028

Will,  C. L.
Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society;

/persons/resource/persons15470

Lührmann,  R.
Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society;

External Ressource
Fulltext (public)

2251100.pdf
(Publisher version), 775KB

Supplementary Material (public)
There is no public supplementary material available
Citation

Schütze, T., Ulrich, A. K. C., Apelt, L., Will, C. L., Bartlick, N., Seeger, M., et al. (2016). Multiple protein-protein interactions converging on the Prp38 protein during activation of the human spliceosome. RNA, 22(2), 265-277. doi:10.1261/rna.054296.115.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0029-C3C3-9
Abstract
Spliceosomal Prp38 proteins contain a conserved amino-terminal domain, but only higher eukaryotic orthologs also harbor a carboxy-terminal RS domain, a hallmark of splicing regulatory SR proteins. We show by crystal structure analysis that the amino-terminal domain of human Prp38 is organized around three pairs of antiparallel alpha-helices and lacks similarities to RNA binding domains found in canonical SR proteins. Instead, yeast two-hybrid analyses suggest that the amino-terminal domain is a versatile protein protein interaction hub that possibly binds 12 other spliceosomal proteins, most of which are recruited at the same stage. as Prp38. By quantitative, alanine surface-scanning two-hybrid screens and biochemical analyses we delineated four distinct interfaces on the Prp38 amino-terminal domain. In vitro interaction assays using recombinant proteins showed that Prp38 can bind at least two proteins simultaneously via two different interfaces. Addition of excess Prp38 amino-terminal domain to in vitro splicing assays, but not of an interaction-deficient mutant, stalled splicing at a precatalytic stage. Our results show that human Prp38 is an unusual SR protein, whose amino-terminal domain is a multi-interface protein protein interaction platform that might organize the relative positioning of other proteins during splicing.