Regulation of constitutive protein transit by phospholipase D in HT29-cl19A cells
Regulation of constitutive protein transit by phospholipase D in HT29-cl19A cells
复制标题
DOI：
10.1074/jbc.m104276200
复制
发表时间：
2001-12-28
影响因子：
4.8
通讯作者：
Raymond, MN
中科院分区：
文献类型：
--
作者：
Denmat-Ouisse, LA;Phebidias, C;Raymond, MN
磷脂酶D（PLD）在控制囊泡出芽和蛋白质转运中起着重要作用。我们先前表明，在静息的上皮HT 29-cl 19 A细胞中，PLD涉及控制组成性蛋白质转运，从trans-Golgi网络到质膜，并且蛋白质转运的佛波醇酯刺激与PLD活化相关（Auger，R.，Robin，P.，Camier，B.，Vial，G.，Rossignol，B.，Tenu，J.- P.，雷蒙德，M。N.（1999）J.Biol.Chem.274，28652-28659）。本研究证明：1）PLD不参与蛋白质转运的早期阶段，2）PLD控制顶侧而非基底侧蛋白质转运，3）HT 29-cl 19 A细胞表达PLD 1b和PLD 2a mRNA和蛋白质，4）HT 29-cl 19 A细胞表达PLD 1b和PLD 2a mRNA和蛋白质，5）HT 29-cl 19 A细胞表达PLD 1b和PLD 2a mRNA和蛋白质，6）HT 29-cl 19 A细胞表达PLD 1b和PLD 2a mRNA和蛋白质。4）PLD 2的催化失活突变体的表达（mPLD 2-K758 R）显著抑制顶端组成性蛋白转运，而PLD 1的催化失活突变体的表达（hPLD 1b-K898 R）阻止了由佛波醇酯触发的顶端转运速率的增加; 5）PLD 2似乎位于包含高尔基体的核周区域，而PLD 1分散在细胞内。静息细胞中的胞质在佛波酯刺激后移位至质膜。两者合计，这些数据导致的结论是，在HT 29 cl 19 A细胞，两个PLD调节蛋白质之间的trans-Golgi网络和顶端质膜，但他们这样做在不同的步骤中的途径。
Phospholipase D (PLD) plays a central role in the control of vesicle budding and protein transit. We previously showed that in resting epithelial HT29-cl19A cells, PLD is implicated in the control of constitutive protein transit, from the trans-Golgi network to the plasma membrane, and that phorbol ester stimulation of protein transit is correlated with PLD activation (Auger, R., Robin, P., Camier, B., Vial, G., Rossignol, B., Tenu, J.-P., and Raymond, M.-N. (1999) J. Biol. Chem. 274, 28652-28659). In this paper we demonstrate that: 1) PLD is not implicated in the earliest phases of protein transit; 2) PLD controls apical but not basolateral protein transit; 3) HT29-cl19A cells express PLD1b and PLD2a mRNAs and proteins; 4) the expression of a catalytically inactive mutant of PLD2 (mPLD2-K758R) significantly inhibited apical constitutive protein transit whereas expression of a catalytically inactive mutant of PLD1 (hPLD1b-K898R) prevented increases in the rate of apical transit as triggered by phorbol esters; 5) PLD2 appears to be located in a perinuclear region containing the Golgi whereas PLD1, which is scattered in the cyto. plasm in resting cells, is translocated to the plasma membrane after phorbol ester stimulation. Taken together, these data lead to the conclusion that in HT29cl19A cells, both PLDs regulate protein transit between the trans-Golgi network and the apical plasma membrane, but that they do so at different steps in the pathway.