Mitotic Golgi vesiculation involves mechanisms independent of Ser25 phosphorylation of GM130.
During mitosis, the Golgi undergoes two sequential fragmentation steps to break from ribbon to individual stacks, then from stacks to vesicles. While the mechanism that regulates the first step has been studied, it remains obscure how the second vesiculation step is regulated. It has been suggested that Cdk1-dependent phosphorylation of the cis-Golgi matrix protein GM130 regulates the second step. Here we have tested if phorphorylation of GM130 by Cdk1 is required for Golgi vesiculation and mitotic progression. Inhibition of Cdk1 activity caused a failure of Golgi vesiculation and defects in chromosome congression/segregation. Expression of non-phosphorylatable mutant of GM130 (GM130S25A) in cells depleted of endogenous GM130 caused no apparent defects in Golgi vesiculation and mitotic progression. Similarly, no apparent defects in Golgi vesiculation and mitotic progression were observed when GM130S25A was expressed in GM130-deficient CHO cells. Our observations suggest that while Cdk1 based phosphorylation is essential for mitotic Golgi vesiculation, mammalian cells could possess redundant, S25 phosphorylation of GM130 independent pathways that ensure Golgi vesiculation and mitotic progression.
Sundaramoorthy
Sriramkumar
S
Cell Biology Program, Temasek Life Sciences Laboratory, National University of Singapore, Singapore, Singapore.
Goh
Justin Bryan Gumagay
JB
Rafee
Shazmina
S
Murata-Hori
Maki
M
D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D002460:Cell Line; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008938:Mitosis; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D001324:Autoantigens; C097741:Golgin subfamily A member 2; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010768:Phosphoserine; D016203:CDC2 Protein Kinase