Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER
The Svp26 protein of S. cerevisiae is an ER- and Golgi-localized integral membrane protein with 4 potential membrane-spanning domains. It functions as an adaptor protein that facilitates the ER exit of Ktr3, a mannosyltransferase required for biosynthesis of O-linked oligosaccharides, and the ER exi...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2014-07-01
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| Series: | Biology Open |
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| Online Access: | http://bio.biologists.org/content/3/3/209 |
| _version_ | 1818461267767066624 |
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| author | Yoichi Noda Takehiro Hara Minako Ishii Koji Yoda |
| author_facet | Yoichi Noda Takehiro Hara Minako Ishii Koji Yoda |
| author_sort | Yoichi Noda |
| collection | DOAJ |
| description | The Svp26 protein of S. cerevisiae is an ER- and Golgi-localized integral membrane protein with 4 potential membrane-spanning domains. It functions as an adaptor protein that facilitates the ER exit of Ktr3, a mannosyltransferase required for biosynthesis of O-linked oligosaccharides, and the ER exit of Mnn2 and Mnn5, mannosyltransferases, which participate in the biosynthesis of N-linked oligosaccharides. Ktr3 belongs to the Kre2 family, which consists of 9 members of type-II membrane proteins sharing sequence similarities. In this report, we examined all Kre2 family members and found that the Golgi localizations of two others, Kre2 and Ktr1, were dependent on Svp26 by immunofluorescence microscopy and cell fractionations in sucrose density gradients. We show that Svp26 functions in facilitating the ER exit of Kre2 and Ktr1 by an in vitro COPII budding assay. Golgi localization of Ktr4 was not dependent on Svp26. Screening null mutants of the genes encoding abundant COPII membrane proteins for those showing mislocalization of Ktr4 in the ER revealed that Erv41 and Erv46 are required for the correct Golgi localization of Ktr4. We provide biochemical evidence that the Erv41-Erv46 complex functions as an adaptor protein for ER exit of Ktr4. This is the first demonstration of the molecular function of this evolutionally conserved protein complex. The domain switching experiments show that the lumenal domain of Ktr4 is responsible for recognition by the Erv41-Erv46 complex. Thus, ER exit of Kre2-family proteins is dependent on distinct adaptor proteins and our results provide new insights into the traffic of Kre2-family mannosyltransferases. |
| first_indexed | 2024-12-14T23:43:26Z |
| format | Article |
| id | doaj.art-f922cb97620941519b253e13076203e2 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-14T23:43:26Z |
| publishDate | 2014-07-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-f922cb97620941519b253e13076203e22022-12-21T22:43:28ZengThe Company of BiologistsBiology Open2046-63902014-07-013320922410.1242/bio.2014631220146312Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ERYoichi NodaTakehiro HaraMinako IshiiKoji YodaThe Svp26 protein of S. cerevisiae is an ER- and Golgi-localized integral membrane protein with 4 potential membrane-spanning domains. It functions as an adaptor protein that facilitates the ER exit of Ktr3, a mannosyltransferase required for biosynthesis of O-linked oligosaccharides, and the ER exit of Mnn2 and Mnn5, mannosyltransferases, which participate in the biosynthesis of N-linked oligosaccharides. Ktr3 belongs to the Kre2 family, which consists of 9 members of type-II membrane proteins sharing sequence similarities. In this report, we examined all Kre2 family members and found that the Golgi localizations of two others, Kre2 and Ktr1, were dependent on Svp26 by immunofluorescence microscopy and cell fractionations in sucrose density gradients. We show that Svp26 functions in facilitating the ER exit of Kre2 and Ktr1 by an in vitro COPII budding assay. Golgi localization of Ktr4 was not dependent on Svp26. Screening null mutants of the genes encoding abundant COPII membrane proteins for those showing mislocalization of Ktr4 in the ER revealed that Erv41 and Erv46 are required for the correct Golgi localization of Ktr4. We provide biochemical evidence that the Erv41-Erv46 complex functions as an adaptor protein for ER exit of Ktr4. This is the first demonstration of the molecular function of this evolutionally conserved protein complex. The domain switching experiments show that the lumenal domain of Ktr4 is responsible for recognition by the Erv41-Erv46 complex. Thus, ER exit of Kre2-family proteins is dependent on distinct adaptor proteins and our results provide new insights into the traffic of Kre2-family mannosyltransferases.http://bio.biologists.org/content/3/3/209GolgiEndoplasmic reticulumCOPII vesicle proteinsErv41Erv46Ktr4Svp26 |
| spellingShingle | Yoichi Noda Takehiro Hara Minako Ishii Koji Yoda Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER Biology Open Golgi Endoplasmic reticulum COPII vesicle proteins Erv41 Erv46 Ktr4 Svp26 |
| title | Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER |
| title_full | Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER |
| title_fullStr | Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER |
| title_full_unstemmed | Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER |
| title_short | Distinct adaptor proteins assist exit of Kre2-family proteins from the yeast ER |
| title_sort | distinct adaptor proteins assist exit of kre2 family proteins from the yeast er |
| topic | Golgi Endoplasmic reticulum COPII vesicle proteins Erv41 Erv46 Ktr4 Svp26 |
| url | http://bio.biologists.org/content/3/3/209 |
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