ABSTRACT
The endoplasmic reticulum (ER) is highly plastic, and increased expression of distinct single ER-resident membrane proteins, such as HMG-CoA reductase (HMGR), can induce a dramatic restructuring of ER membranes into highly organized arrays. Studies on the ER-remodeling behavior of the two yeast HMGR isozymes, Hmg1p and Hmg2p, suggest that they could be mechanistically distinct. We examined the features of Hmg2p required to generate its characteristic structures, and we found that the molecular requirements are similar to those of Hmg1p. However, the structures generated by Hmg1p and Hmg2p have distinct cell biological features determined by the transmembrane regions of the proteins. In parallel, we conducted a genetic screen to identify HER genes (required for Hmg2p-induced ER Remodeling), further confirming that the mechanisms of membrane reorganization by these two proteins are distinct because most of the HER genes were required for Hmg2p but not Hmg1p-induced ER remodeling. One of the HER genes identified was PSD1, which encodes the phospholipid biosynthetic enzyme phosphatidylserine decarboxylase. This direct connection to phospholipid biosynthesis prompted a more detailed examination of the effects of Hmg2p on phospholipid mutants and composition. Our analysis revealed that overexpression of Hmg2p caused significant and specific growth defects in nulls of the methylation pathway for phosphatidylcholine biosynthesis that includes the Psd1p enzyme. Furthermore, increased expression of Hmg2p altered the composition of cellular phospholipids in a manner that implied a role for PSD1. These phospholipid effects, unlike Hmg2p-induced ER remodeling, required the enzymatic activity of Hmg2p. Together, our results indicate that, although related, Hmg2p- and Hmg1p-induced ER remodeling are mechanistically distinct.
Subject(s)
Endoplasmic Reticulum/metabolism , Gene Expression Regulation, Fungal , HMGB1 Protein/genetics , HMGB2 Protein/chemistry , HMGB2 Protein/genetics , Catalysis , Cell Membrane/metabolism , Cell Proliferation , Fungal Proteins/metabolism , HMGB1 Protein/chemistry , Microscopy, Fluorescence/methods , Models, Biological , Models, Genetic , Phospholipids/chemistry , Phospholipids/metabolism , Protein Isoforms , Protein Structure, Tertiary , Recombinant Proteins/chemistryABSTRACT
The unfolded protein response (UPR) pathway helps cells cope with endoplasmic reticulum (ER) stress by activating genes that increase the ER's functional capabilities. We have identified a novel role for the UPR pathway in facilitating budding yeast cytokinesis. Although other cell cycle events are unaffected by conditions that disrupt ER function, cytokinesis is sensitive to these conditions. Moreover, efficient cytokinesis requires the UPR pathway even during unstressed growth conditions. UPR-deficient cells are defective in cytokinesis, and cytokinesis mutants activate the UPR. The UPR likely achieves its role in cytokinesis by sensing small changes in ER load and making according changes in ER capacity. We propose that cytokinesis is one of many cellular events that require a subtle increase in ER function and that the UPR pathway has a previously uncharacterized housekeeping role in maintaining ER plasticity during normal cell growth.
Subject(s)
Cytokinesis , Endoplasmic Reticulum/metabolism , Cell Cycle , Endoplasmic Reticulum/genetics , Molecular Chaperones , Protein Folding , SaccharomycetalesABSTRACT
The endoplasmic reticulum (ER) is an extremely plastic and dynamic organelle. Its size and shape can undergo drastic changes to meet changing demands for ER-related functions, or as a response to drugs or pathogens. Because of the ER's key functions in protein and lipid synthesis, this organelle is a hotbed of detailed molecular analysis.
