Abstract
Ubiquitination of proteins modifies protein function by either altering their activities, promoting their degradation, or altering their subcellular localization. Deubiquitinating enzymes are proteases that reverse this ubiquitination. Previous studies demonstrate that proteins that contain an ovarian tumor (OTU) domain possess deubiquitinating activity. This domain of ∼130 amino acids is weakly similar to the papain family of proteases and is highly conserved from yeast to mammals. Here we report structural and functional studies on the OTU domain-containing protein from yeast, Otu1. We show that Otu1 binds polyubiquitin chain analogs more tightly than monoubiquitin and preferentially hydrolyzes longer polyubiquitin chains with Lys48 linkages, having little or no activity on Lys63- and Lys 29-linked chains. We also show that Otu1 interacts with Cdc48, a regulator of the ER-associated degradation pathway. We also report the x-ray crystal structure of the OTU domain of Otu1 covalently complexed with ubiquitin and carry out structure-guided mutagenesis revealing a novel mode of ubiquitin recognition and a variation on the papain protease catalytic site configuration that appears to be conserved within the OTU family of ubiquitin hydrolases. Together, these studies provide new insights into ubiquitin binding and hydrolysis by yeast Otu1 and other OTU domain-containing proteins.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 11038-11049 |
| Number of pages | 12 |
| Journal | Journal of Biological Chemistry |
| Volume | 283 |
| Issue number | 16 |
| DOIs | |
| State | Published - Apr 18 2008 |
| Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Cell Biology
- Molecular Biology
Cite this
Structural basis for ubiquitin recognition by the Otu1 ovarian tumor domain protein. / Messick, Troy Eugene; Russell, Nathaniel Scott; Iwata, Ayaka Jennifer; Sarachan, Kathryn Lorenz; Shiekhattar, Ramin; Shanks, John R.; Reyes-Turcu, Francisca E.; Wilkinson, Keith D.; Marmorstein, Ronen.
In: Journal of Biological Chemistry, Vol. 283, No. 16, 18.04.2008, p. 11038-11049.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Structural basis for ubiquitin recognition by the Otu1 ovarian tumor domain protein
AU - Messick, Troy Eugene
AU - Russell, Nathaniel Scott
AU - Iwata, Ayaka Jennifer
AU - Sarachan, Kathryn Lorenz
AU - Shiekhattar, Ramin
AU - Shanks, John R.
AU - Reyes-Turcu, Francisca E.
AU - Wilkinson, Keith D.
AU - Marmorstein, Ronen
PY - 2008/4/18
Y1 - 2008/4/18
N2 - Ubiquitination of proteins modifies protein function by either altering their activities, promoting their degradation, or altering their subcellular localization. Deubiquitinating enzymes are proteases that reverse this ubiquitination. Previous studies demonstrate that proteins that contain an ovarian tumor (OTU) domain possess deubiquitinating activity. This domain of ∼130 amino acids is weakly similar to the papain family of proteases and is highly conserved from yeast to mammals. Here we report structural and functional studies on the OTU domain-containing protein from yeast, Otu1. We show that Otu1 binds polyubiquitin chain analogs more tightly than monoubiquitin and preferentially hydrolyzes longer polyubiquitin chains with Lys48 linkages, having little or no activity on Lys63- and Lys 29-linked chains. We also show that Otu1 interacts with Cdc48, a regulator of the ER-associated degradation pathway. We also report the x-ray crystal structure of the OTU domain of Otu1 covalently complexed with ubiquitin and carry out structure-guided mutagenesis revealing a novel mode of ubiquitin recognition and a variation on the papain protease catalytic site configuration that appears to be conserved within the OTU family of ubiquitin hydrolases. Together, these studies provide new insights into ubiquitin binding and hydrolysis by yeast Otu1 and other OTU domain-containing proteins.
AB - Ubiquitination of proteins modifies protein function by either altering their activities, promoting their degradation, or altering their subcellular localization. Deubiquitinating enzymes are proteases that reverse this ubiquitination. Previous studies demonstrate that proteins that contain an ovarian tumor (OTU) domain possess deubiquitinating activity. This domain of ∼130 amino acids is weakly similar to the papain family of proteases and is highly conserved from yeast to mammals. Here we report structural and functional studies on the OTU domain-containing protein from yeast, Otu1. We show that Otu1 binds polyubiquitin chain analogs more tightly than monoubiquitin and preferentially hydrolyzes longer polyubiquitin chains with Lys48 linkages, having little or no activity on Lys63- and Lys 29-linked chains. We also show that Otu1 interacts with Cdc48, a regulator of the ER-associated degradation pathway. We also report the x-ray crystal structure of the OTU domain of Otu1 covalently complexed with ubiquitin and carry out structure-guided mutagenesis revealing a novel mode of ubiquitin recognition and a variation on the papain protease catalytic site configuration that appears to be conserved within the OTU family of ubiquitin hydrolases. Together, these studies provide new insights into ubiquitin binding and hydrolysis by yeast Otu1 and other OTU domain-containing proteins.
UR - http://www.scopus.com/inward/record.url?scp=44849136272&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=44849136272&partnerID=8YFLogxK
U2 - 10.1074/jbc.M704398200
DO - 10.1074/jbc.M704398200
M3 - Article
VL - 283
SP - 11038
EP - 11049
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 16
ER -