English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  The Prp8 RNase H-like domain inhibits Brr2-mediated U4/U6 snRNA unwinding by blocking Brr2 loading onto the U4 snRNA.

Mozaffari-Jovin, S., Santos, K. F., Hsiao, H. H., Will, C. L., Urlaub, H., Wahl, M. C., et al. (2012). The Prp8 RNase H-like domain inhibits Brr2-mediated U4/U6 snRNA unwinding by blocking Brr2 loading onto the U4 snRNA. Genes and Development, 26(23), 2422-2434. doi:10.1101/gad.200949.112.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-25FA-F Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-C163-2
Genre: Journal Article

Files

show Files
hide Files
:
1570032.pdf (Publisher version), 947KB
Name:
1570032.pdf
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/pdf / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-
:
1570032-Suppl.doc (Supplementary material), 9MB
Name:
1570032-Suppl.doc
Description:
-
Visibility:
Public
MIME-Type / Checksum:
application/msword / [MD5]
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show
hide
Description:
-

Creators

show
hide
 Creators:
Mozaffari-Jovin, S.1, Author              
Santos, K. F., Author
Hsiao, H. H.2, Author              
Will, C. L.1, Author              
Urlaub, H.2, Author              
Wahl, M. C., Author
Lührmann, R.1, Author              
Affiliations:
1Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              
2Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              

Content

show
hide
Free keywords: Pre-mRNA splicing; RNA helicase; RNA–protein complex; RNA–protein cross-linking; spliceosome catalytic activation
 Abstract: The spliceosomal RNA helicase Brr2 catalyzes unwinding of the U4/U6 snRNA duplex, an essential step for spliceosome catalytic activation. Brr2 is regulated in part by the spliceosomal Prp8 protein by an unknown mechanism. We demonstrate that the RNase H (RH) domain of yeast Prp8 binds U4/U6 small nuclear RNA (snRNA) with the single-stranded regions of U4 and U6 preceding U4/U6 stem I, contributing to its binding. Via cross-linking coupled with mass spectrometry, we identify RH domain residues that contact the U4/U6 snRNA. We further demonstrate that the same single-stranded region of U4 preceding U4/U6 stem I is recognized by Brr2, indicating that it translocates along U4 and first unwinds stem I of the U4/U6 duplex. Finally, we show that the RH domain of Prp8 interferes with U4/U6 unwinding by blocking Brr2's interaction with the U4 snRNA. Our data reveal a novel mechanism whereby Prp8 negatively regulates Brr2 and potentially prevents premature U4/U6 unwinding during splicing. They also support the idea that the RH domain acts as a platform for the exchange of U6 snRNA for U1 at the 5′ splice site. Our results provide insights into the mechanism whereby Brr2 unwinds U4/U6 and show how this activity is potentially regulated prior to spliceosome activation.

Details

show
hide
Language(s): eng - English
 Dates: 20122012-12-01
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1101/gad.200949.112
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Genes and Development
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 26 (23) Sequence Number: - Start / End Page: 2422 - 2434 Identifier: -