role of human sec63 in modulating the steady-state levels of multi-spanning membrane proteins人类sec63在调制多跨膜蛋白的稳态水平.pdfVIP

Role of Human Sec63 in Modulating the Steady-State Levels of Multi-Spanning Membrane Proteins 1. 2. 2 2 ¨ 2 ¨ 2 Andreas Mades , Katherina Gotthardt , Karin Awe , Jens Stieler , Tatjana Doring , Sabine Fuser , Reinhild Prange2* 1 Department of Medicine III, Hematology and Oncology, Johannes Gutenberg-University School of Medicine, Mainz, Germany, 2 Department of Medical Microbiology and Hygiene, University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany Abstract The Sec61 translocon of the endoplasmic reticulum (ER) membrane forms an aqueous pore, allowing polypeptides to be transferred across or integrated into membranes. Protein translocation into the ER can occur co- and posttranslationally. In yeast, posttranslational translocation involves the heptameric translocase complex including its Sec62p and Sec63p subunits. The mammalian ER membrane contains orthologs of yeast Sec62p and Sec63p, but their function is poorly understood. Here, we analyzed the effects of excess and deficit Sec63 on various ER cargoes using human cell culture systems. The overexpression of Sec63 reduces the steady-state levels of viral and cellular multi-spanning membrane proteins in a cotranslational mode, while soluble and single-spanning ER reporters are not affected. Consistent with this, the knock-down of Sec63 increases the steady-state pools of polytopic ER proteins, suggesting a substrate-specific and regulatory function of Sec63 in ER import. Overexpressed Sec63 exerts its down-regulating activity on polytopic protein levels independent of its Sec62-interacting motif, indicating that it may not