Two distinct Vps34 phosphatidylinositol 3-kinase complexes function in autophagy and carboxypeptidase Y sorting in Saccharomyces cerevisiae.
Two distinct Vps34 phosphatidylinositol 3-kinase complexes function in autophagy and carboxypeptidase Y sorting in Saccharomyces cerevisiae.
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DOI：
10.1083/jcb.152.3.519
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发表时间：
2001-02-05
期刊：
影响因子：
--
通讯作者：
Ohsumi Y
中科院分区：
文献类型：
--
作者：
Kihara A;Noda T;Ishihara N;Ohsumi Y
Vps 30 p/Apg 6p是自噬和羧肽酶Y（CPY）分选所必需的。虽然已知Vps 30 p与Apg 14 p相互作用，但其确切作用仍不清楚。我们发现有两个蛋白与Vps 30 p共纯化。经质谱鉴定为Vps 38 p和Vps 34 p，一种磷脂酰肌醇（PtdIns）3-激酶。将Vps 34 p、Vps 38 p、Apg 14 p和Vps 34 p的激活剂Vps 15 p与Vps 30 p共免疫沉淀。这些结果表明，Vps 30 p作为Vps 34 PtdIns 3-激酶复合物的亚基起作用。表型分析表明Apg 14 p和Vps 38 p分别是自噬和CPY分选所需的，而Vps 30 p、Vps 34 p和Vps 15 p是这两个过程所需的。使用抗Apg 14 p和抗Vps 38 p抗体的免疫共沉淀和下拉实验表明，存在两种不同的Vps 34 PtdIns 3-激酶复合物：一种含有Vps 15 p、Vps 30 p和Apg 14 p，在自噬中起作用，另一种含有Vps 15 p、Vps 30 p和Vps 38 p，在CPY分选中起作用。vps 34和vps 15突变体显示了额外的表型，如蛋白酶A和蛋白酶B转运缺陷，这意味着存在另一种PtdIns 3-激酶复合物。我们提出，多个Vps 34 p-Vps 15 p复合物与特定的调节蛋白可能会履行其膜运输事件在不同的网站。
Vps30p/Apg6p is required for both autophagy and sorting of carboxypeptidase Y (CPY). Although Vps30p is known to interact with Apg14p, its precise role remains unclear. We found that two proteins copurify with Vps30p. They were identified by mass spectrometry to be Vps38p and Vps34p, a phosphatidylinositol (PtdIns) 3–kinase. Vps34p, Vps38p, Apg14p, and Vps15p, an activator of Vps34p, were coimmunoprecipitated with Vps30p. These results indicate that Vps30p functions as a subunit of a Vps34 PtdIns 3–kinase complex(es). Phenotypic analyses indicated that Apg14p and Vps38p are each required for autophagy and CPY sorting, respectively, whereas Vps30p, Vps34p, and Vps15p are required for both processes. Coimmunoprecipitation using anti-Apg14p and anti-Vps38p antibodies and pull-down experiments showed that two distinct Vps34 PtdIns 3–kinase complexes exist: one, containing Vps15p, Vps30p, and Apg14p, functions in autophagy and the other containing Vps15p, Vps30p, and Vps38p functions in CPY sorting. The vps34 and vps15 mutants displayed additional phenotypes such as defects in transport of proteinase A and proteinase B, implying the existence of another PtdIns 3–kinase complex(es). We propose that multiple Vps34p–Vps15p complexes associated with specific regulatory proteins might fulfill their membrane trafficking events at different sites.
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影响因子：
7.8
作者：
Baba, M;Osumi, M;Scott, SV;Klionsky, DJ;Ohsumi, Y
通讯作者：
Ohsumi, Y
DOI：
10.1006/bbrc.1995.1636
发表时间：
1995-05-05
影响因子：
3.1
作者：
NODA, T;MATSUURA, A;OHSUMI, Y
通讯作者：
OHSUMI, Y
影响因子：
4.8
作者：
Gaffet, P;Jones, AT;Clague, MJ
通讯作者：
Clague, MJ
影响因子：
4.8
作者：
Noda, T;Ohsumi, Y
通讯作者：
Ohsumi, Y
影响因子：
5.3
作者：
IRIE, K;TAKASE, M;OSHIMA, Y
通讯作者：
OSHIMA, Y