Membrane targeting of p115 phosphorylation mutants and their effects on Golgi integrity and secretory traffic.
The cytosolic phosphoprotein p115 is required for ER to Golgi traffic and for Golgi reassembly after mitosis. In cells, p115 is localized to ER exit sites, ER-Golgi Intermediate Compartment (ERGIC) and the Golgi, and cycles between these compartments. P115 is phosphorylated on serine 942, and this modification appears to control p115 association with membranes. P115 is likely to function by reversibly interacting with effector proteins, and in the Golgi, two proteins, GM130 and giantin, have been shown to bind p115. The GM130-p115 and the giantin-p115 interactions are enhanced by p115 phosphorylation. Phosphorylation appears to be essential for p115 function, since substitutions of serine 942 abolish p115 ability to sustain cisternal reformation in an in vitro assay reconstituting Golgi reassembly after mitosis. Here, we explored how phosphorylation of p115 affects its intracellular targeting to distinct cellular compartments, and its function in secretory traffic. We generated phosphorylation mutants of p115 and tested their ability to target to ER exit sites, ERGIC and the Golgi. In addition, we explored whether expression of the mutants causes disruption of Golgi structure and perturbs ER-Golgi traffic of a VSV-G cargo protein.
Brandon
Elizabeth
E
Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
Gao
Yasheng
Y
Garcia-Mata
Rafael
R
Alvarez
Cecilia
C
Sztul
Elizabeth
E
D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002451:Cell Compartmentation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D008099:Liver; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D021381:Protein Transport; D051381:Rats; D033921:Vesicular Transport Proteins
D002352:Carrier Proteins; D000077614:Golgi Matrix Proteins; D008565:Membrane Proteins; D033921:Vesicular Transport Proteins; C091580:vesicular transport factor p115