Involvement of Golgi-associated Lyn tyrosine kinase in the translocation of annexin II to the endoplasmic reticulum under oxidative stress.
Src-family tyrosine kinases, known to participate in signaling pathways of a variety of receptors at the plasma membrane, are found in cellular endomembranes such as the Golgi apparatus and endosomes. Recently, we showed that Lyn, a member of the Src kinases, accumulates on the Golgi apparatus and then traffics to the plasma membrane. We show here that a majority of endogenous Lyn but not c-Src is accumulated in Golgi-enriched heavy-membrane fractions on a sucrose-density gradient, whereas a small amount of endogenous Lyn is present in light-membrane fractions containing the plasma membrane. Inducible expression of kinase-active Lyn, which biosynthetically reaches the Golgi apparatus, triggers tyrosine phosphorylation of proteins including annexin II. Coimmunoprecipitation analyses reveal that Lyn physically associates with annexin II, and an in vitro kinase assay shows that Lyn phosphorylates annexin II directly. Furthermore, stimulation of cells with H2O2 induces tyrosine phosphorylation of annexin II on the Golgi apparatus in a manner that is dependent on the kinase activity of Src kinases, leading to the translocation of annexin II from the Golgi apparatus to the endoplasmic reticulum. Thus, these results suggest that endomembranes containing the Golgi apparatus where Lyn is anchored can serve as a signaling platform under oxidative stress.
Matsuda
Daisuke
D
Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, Inohana 1-8-1, Chuo-ku, Chiba 260-8675, Japan.
Nakayama
Yuji
Y
Horimoto
Shinya
S
Kuga
Takahisa
T
Ikeda
Kikuko
K
Kasahara
Kousuke
K
Yamaguchi
Naoto
N
D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000201:enzymology / Q000378:metabolism*; D006056:Golgi Apparatus / Q000201:enzymology*; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D051379:Mice; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology*; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000502:physiology*
D017306:Annexin A2; D016877:Oxidants; D014443:Tyrosine; D006861:Hydrogen Peroxide; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases