HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 8, 4646-4653, February 24, 2006

This Article Full Text Full Text (PDF) Supplmental Data All Versions of this Article: 281/8/4646    most recent M512215200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kreft, S. G. Articles by Hochstrasser, M. Search for Related Content PubMed PubMed Citation Articles by Kreft, S. G. Articles by Hochstrasser, M. Social Bookmarking                      What's this?

Membrane Topology of the Yeast Endoplasmic Reticulum-localized Ubiquitin Ligase Doa10 and Comparison with Its Human Ortholog TEB4 (MARCH-VI)^*

From the Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520-8114

Quality control machinery in the endoplasmic reticulum (ER) helps ensure that only properly folded and assembled proteins accumulate in the ER or continue along the secretory pathway. Aberrant proteins are retrotranslocated to the cytosol and degraded by the proteasome, a process called ER-associated degradation. Doa10, a transmembrane protein of the ER/nuclear envelope, is one of the primary ubiquitin ligases (E3s) participating in ER-associated degradation in Saccharomyces cerevisiae. Here we report the membrane organization of the 1319-residue Doa10 polypeptide. The topology was determined by fusing a dual-topology reporter after 16 different Doa10 fragments. Our results indicate that Doa10 contains 14 transmembrane helices (TMs). Based on protease digestion of yeast microsomes, both the N-terminal RING-CH domain and the C terminus face the cytosol. Notably, the experimentally derived topology was not predicted correctly by any of the generally available TM prediction algorithms. Bioinformatic analysis and in silico mutagenesis guided the topological studies through problematic regions. The conserved TD domain in Doa10 includes three TMs. These TMs might function in cofactor binding or substrate recognition, or they might be part of a retrotranslocation channel. The Derlins were previously proposed to provide such channels, but we show that the two yeast Derlins are not required for degradation of Doa10 membrane substrates, as was found before for the Sec61 translocon. Finally, we provide evidence that the likely human Doa10 ortholog, TEB4 (MARCH-VI), adopts a topology similar to that of Doa10.

Received for publication, November 14, 2005 , and in revised form, December 21, 2005.

^* This work was supported by National Institutes of Health Grant GM46904 (to M. H.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The online version of this article (available at http://www.jbc.org) contains supplemental Figs. S1 and S2.

¹ To whom correspondence should be addressed: Dept. of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Ave., New Haven, CT 06520-8114. E-mail: mark.hochstrasser{at}yale.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. M. Garza, B. K. Sato, and R. Y. Hampton In Vitro Analysis of Hrd1p-mediated Retrotranslocation of Its Multispanning Membrane Substrate 3-Hydroxy-3-methylglutaryl (HMG)-CoA Reductase J. Biol. Chem., May 29, 2009; 284(22): 14710 - 14722. [Abstract] [Full Text] [PDF]

				K. Oresic, C. L. Ng, and D. Tortorella TRAM1 Participates in Human Cytomegalovirus US2- and US11-mediated Dislocation of an Endoplasmic Reticulum Membrane Glycoprotein J. Biol. Chem., February 27, 2009; 284(9): 5905 - 5914. [Abstract] [Full Text] [PDF]

				B. Gereben, A. M. Zavacki, S. Ribich, B. W. Kim, S. A. Huang, W. S. Simonides, A. Zeold, and A. C. Bianco Cellular and Molecular Basis of Deiodinase-Regulated Thyroid Hormone Signaling Endocr. Rev., December 1, 2008; 29(7): 898 - 938. [Abstract] [Full Text] [PDF]

				J. Barrowman, C. Hamblet, C. M. George, and S. Michaelis Analysis of Prelamin A Biogenesis Reveals the Nucleus to be a CaaX Processing Compartment Mol. Biol. Cell, December 1, 2008; 19(12): 5398 - 5408. [Abstract] [Full Text] [PDF]

				D. C. Scott and R. Schekman Role of Sec61p in the ER-associated degradation of short-lived transmembrane proteins J. Cell Biol., October 22, 2008; 181(7): 1095 - 1105. [Abstract] [Full Text] [PDF]

				Y. Xie, O. Kerscher, M. B. Kroetz, H. F. McConchie, P. Sung, and M. Hochstrasser The Yeast Hex3{middle dot}Slx8 Heterodimer Is a Ubiquitin Ligase Stimulated by Substrate Sumoylation J. Biol. Chem., November 23, 2007; 282(47): 34176 - 34184. [Abstract] [Full Text] [PDF]

				Y. Sasagawa, K. Yamanaka, and T. Ogura ER E3 ubiquitin ligase HRD-1 and its specific partner chaperone BiP play important roles in ERAD and developmental growth in Caenorhabditis elegans Genes Cells, September 1, 2007; 12(9): 1063 - 1073. [Abstract] [Full Text] [PDF]

				X. Wang, R. A. Herr, W.-J. Chua, L. Lybarger, E. J.H.J. Wiertz, and T. H. Hansen Ubiquitination of serine, threonine, or lysine residues on the cytoplasmic tail can induce ERAD of MHC-I by viral E3 ligase mK3 J. Cell Biol., May 21, 2007; 177(4): 613 - 624. [Abstract] [Full Text] [PDF]

				M. Lerner, M. Corcoran, D. Cepeda, M. L. Nielsen, R. Zubarev, F. Ponten, M. Uhlen, S. Hober, D. Grander, and O. Sangfelt The RBCC Gene RFP2 (Leu5) Encodes a Novel Transmembrane E3 Ubiquitin Ligase Involved in ERAD Mol. Biol. Cell, May 1, 2007; 18(5): 1670 - 1682. [Abstract] [Full Text] [PDF]

				A. Zeold, L. Pormuller, M. Dentice, J. W. Harney, C. Curcio-Morelli, S. M. Tente, A. C. Bianco, and B. Gereben Metabolic Instability of Type 2 Deiodinase Is Transferable To Stable Proteins Independently of Subcellular Localization J. Biol. Chem., October 20, 2006; 281(42): 31538 - 31543. [Abstract] [Full Text] [PDF]

				G. C. Hassink, M. T. Barel, S. B. Van Voorden, M. Kikkert, and E. J. Wiertz Ubiquitination of MHC Class I Heavy Chains Is Essential for Dislocation by Human Cytomegalovirus-encoded US2 but Not US11 J. Biol. Chem., October 6, 2006; 281(40): 30063 - 30071. [Abstract] [Full Text] [PDF]