Skip Navigation



Nucleic Acids Research Advance Access published online on February 16, 2008
Nucleic Acids Research, doi:10.1093/nar/gkm1152
This Article
Right arrow Full Text Freely available
Right arrow Print PDF (2778K) Freely available
Right arrow Screen PDF (576K) Freely available
Right arrowOA All Versions of this Article:
36/7/2182    most recent
gkm1152v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Ehmsen, K. T.
Right arrow Articles by Heyer, W.-D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ehmsen, K. T.
Right arrow Articles by Heyer, W.-D.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nucleic Acid Enzymes

Saccharomyces cerevisiae Mus81-Mms4 is a catalytic, DNA structure-selective endonuclease

Kirk Tevebaugh Ehmsen1 and Wolf-Dietrich Heyer1,2,*

1Section of Microbiology and 2Section of Molecular and Cellular Biology, University of California Davis, Davis, CA 95616-8665, USA

*To whom correspondence should be addressed. Tel: 530-752-3001; Fax: 530-752-3011; Email: wdheyer{at}ucdavis.edu

Received October 2, 2007. Revised December 7, 2007. Accepted December 12, 2007.

The DNA structure-selective endonuclease Mus81-Mms4/Eme1 is a context-specific recombination factor that supports DNA replication, but is not essential for DSB repair in Saccharomyces cerevisiae. We overexpressed Mus81-Mms4 in S. cerevisiae, purified the heterodimer to apparent homogeneity, and performed a classical enzymological characterization. Kinetic analysis (kcat, KM) demonstrated that Mus81-Mms4 is catalytically active and identified three substrate classes in vitro. Class I substrates reflect low KM (3–7 nM) and high kcat (~1 min–1) and include the nicked Holliday junction, 3'-flapped and replication fork-like structures. Class II substrates share low KM (1–6 nM) but low kcat (≤0.3 min–1) relative to Class I substrates and include the D-loop and partial Holliday junction. The splayed Y junction defines a class III substrate having high KM (~30 nM) and low kcat (0.26 min–1). Holliday junctions assembled from oligonucleotides with or without a branch migratable core were negligibly cut in vitro. We found that Mus81 and Mms4 are phosphorylated constitutively and in the presence of the genotoxin MMS. The endogenous complex purified in either modification state is negligibly active on Holliday junctions. Hence, Holliday junction incision activity in vitro cannot be attributed to the Mus81-Mms4 heterodimer in isolation.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Plant Physiol.Home page
V. Geuting, D. Kobbe, F. Hartung, J. Durr, M. Focke, and H. Puchta
Two Distinct MUS81-EME1 Complexes from Arabidopsis Process Holliday Junctions
Plant Physiology, June 1, 2009; 150(2): 1062 - 1071.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
K. T. Ehmsen and W.-D. Heyer
A junction branch point adjacent to a DNA backbone nick directs substrate cleavage by Saccharomyces cerevisiae Mus81-Mms4
Nucleic Acids Res., April 1, 2009; 37(6): 2026 - 2036.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Matulova, V. Marini, R. C. Burgess, A. Sisakova, Y. Kwon, R. Rothstein, P. Sung, and L. Krejci
Cooperativity of Mus81{middle dot}Mms4 with Rad54 in the Resolution of Recombination and Replication Intermediates
J. Biol. Chem., March 20, 2009; 284(12): 7733 - 7745.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
O. M. Mazina and A. V. Mazin
Human Rad54 protein stimulates human Mus81-Eme1 endonuclease
PNAS, November 25, 2008; 105(47): 18249 - 18254.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.