Activation of Lyn Tyrosine Kinase through Decreased Membrane Cholesterol Levels during a Change in Its Membrane Distribution upon Cell Detachment

Morinaga, Takao; Abe, Kohei; Nakayama, Yuji; Yamaguchi, Noritaka; Yamaguchi, Naoto

doi:10.1074/jbc.m114.580001

Cited by 10 publications

(30 citation statements)

References 53 publications

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“…3) We showed that newly synthesized Lyn, a member of Src-family kinases, in the cytoplasm is biosynthetically accumulated at the Golgi, which contains various membranous cisternae and clusters of vesicles, including cis-, medial-and trans-Golgi cisternae and the trans-Golgi network (TGN), 4,5) and Golgi-associated Lyn is subsequently transported to the plasma membrane. [6][7][8][9][10] Since the localization of proteins is linked to their functions, we also showed that Golgi-associated Lyn can serve as a signaling platform under oxidative stress. 7) Although newly synthesized secretory proteins move through the secretory pathway: rough endoplasmic reticulum (ER) cis-, medial-and trans-Golgi cisternae secretory or transport vesicles plasma membrane, the intra-Golgi localization of Lyn remains elusive.…”

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confidence: 93%

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Golgi Distribution of Lyn to Caveolin- and Giantin-Positive cis-Golgi Membranes and the Caveolin-Negative, TGN46-Positive trans-Golgi Network

Okamoto

Morinaga

Yamaguchi

et al. 2018

Biological & Pharmaceutical Bulletin

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Src-family tyrosine kinases, classified as cytosolic enzymes, have crucial roles in regulating cell proliferation, differentiation, migration and cell-shape changes. Newly synthesized Lyn, a member of Src-family kinases, is biosynthetically accumulated at the cytoplasmic face of caveolin-containing Golgi membranes via posttranslational lipid modifications and then transported to the plasma membrane. However, the precise intra-Golgi localization of Lyn remains elusive. By means of a 19°C block-release technique and short-term brefeldin A treatment, we show here that the distribution of Lyn is not monotonously spread within the Golgi but selectively intensified in two distinct membrane compartments: giantin-and caveolin-positive membranes and trans-Golgi network protein (TGN)46-positive but caveolin-negative membranes. Furthermore, Lyn exits the Golgi from the caveolin-positive cis-Golgi cisternae or the caveolin-negative trans-Golgi network. These results suggest that Lyn moves apart from caveolin, a secretory protein, within the Golgi during Lyn's trafficking to the plasma membrane.

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confidence: 93%

“…10,15) Confocal images were obtained using a Fluoview FV500 (Olympus, Japan) laser scanning microscope with a 60× water-immersion objective. The Pearson's R values were determined as described.…”

Section: )mentioning

confidence: 99%

Golgi Distribution of Lyn to Caveolin- and Giantin-Positive cis-Golgi Membranes and the Caveolin-Negative, TGN46-Positive trans-Golgi Network

Okamoto

Morinaga

Yamaguchi

et al. 2018

Biological & Pharmaceutical Bulletin

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“…Knockdown of c-Src was performed with shRNA for silencing c-Src (sh(c-Src); 5Ј-GCTCCAGATTGTCAACAAC-3Ј) (68). Knockdown of Fyn was performed with shRNA for silencing Fyn (sh(Fyn); 5Ј-GGAAGAGCTCTGAAATTAC-3Ј) (69). Knockdown of Lyn was performed with shRNA for silencing Lyn (sh(Lyn); 5Ј-GGACAGAGGTTTCAAACTA-3Ј) (69).…”

Section: Methodsmentioning

confidence: 99%

“…HeLa S3/TR cells, which can express v-Src (HeLa S3/TR/v-Src) or c-Src-HA, were prepared recently (79,80). HeLa S3/sh(Fyn) cells, HeLa S3/sh(Lyn) cells, and HeLa S3/sh(Fyn)ϩsh(Lyn) cells were generated by using pENTR4/H1/sh(Fyn) and/or pENTR4/H1/sh(Lyn) with a plasmid containing the hygromycin resistant gene as described previously (69). Cells seeded in a 35-mm (60-mm) culture dish were transiently transfected with 1 g (3 g) of plasmid DNA using 5 g (15 g) of linear polyethyleneimine (25 kDa) (Polyscience, Inc.) (78).…”

Section: Methodsmentioning

confidence: 99%

“…Knockdown of Fyn was performed with shRNA for silencing Fyn (sh(Fyn); 5Ј-GGAAGAGCTCTGAAATTAC-3Ј) (69). Knockdown of Lyn was performed with shRNA for silencing Lyn (sh(Lyn); 5Ј-GGACAGAGGTTTCAAACTA-3Ј) (69). The nucleotides for shRNA were annealed and subcloned into the BglII-XbaI site of the pENTR4/H1 vector (provided by H. Miyoshi) (19, 70 -72).…”

Section: Methodsmentioning

confidence: 99%

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Src Acts as an Effector for Ku70-dependent Suppression of Apoptosis through Phosphorylation of Ku70 at Tyr-530

Morii

Kubota

Honda

et al. 2017

Journal of Biological Chemistry

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Edited by Xiao-Fan WangSrc-family tyrosine kinases are widely expressed in many cell types and participate in a variety of signal transduction pathways. Despite the significance of Src in suppression of apoptosis, its mechanism remains poorly understood. Here we show that Src acts as an effector for Ku70-dependent suppression of apoptosis. Inhibition of endogenous Src activity promotes UV-induced apoptosis, which is impaired by Ku70 knockdown. Src phosphorylates Ku70 at Tyr-530, being close to the possible acetylation sites involved in promotion of apoptosis. Src-mediated phosphorylation of Ku70 at Tyr-530 decreases acetylation of Ku70, whereas Src inhibition augments acetylation of Ku70. Importantly, knockdown-rescue experiments with stable Ku70 knockdown cells show that the nonphosphorylatable Y530F mutant of Ku70 reduces the ability of Ku70 to suppress apoptosis accompanied by augmentation of Ku70 acetylation. Our results reveal that Src plays a protective role against hyperactive apoptotic cell death by reducing apoptotic susceptibility through phosphorylation of Ku70 at Tyr-530.

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Fyn Accelerates M Phase Progression by Promoting the Assembly of Mitotic Spindle Microtubules

Okamoto

Nakayama

Kakihana

et al. 2015

J of Cellular Biochemistry

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The mitotic spindle is the major piece of cellular machinery essential for faithful chromosome segregation. Whereas Fyn, a member of Src-family kinases, is known to be localized to the meiotic and mitotic spindle microtubules, the role of Fyn in mitotic spindle formation has not yet been completely elucidated. In this study, we studied the role of Fyn in spindle formation and effects on M-phase progression. Re-expression of Fyn induced increases in the fluorescence intensity of mitotic spindle microtubules in SYF cells having triple knock-out mutations of c-Src, c-Yes, and Fyn. Cold treatment results showed that Fyn increases the maximum length of microtubules in HeLa S3 cells in a manner dependent on Fyn kinase activity. Complete depolymerization of microtubules under cold treatment and the following release into 37 °C revealed that the increase in the microtubule length in Fyn-expressing cells may be attributed to the promotion of microtubule polymerization. After cold treatment, Fyn promotes the accumulation of EB1, which is a plus-end tracking protein and facilitates microtubule growth, in a manner dependent on the kinase activity. Furthermore, Fyn accelerates the M phase progression of cells from nocodazole arrest. These results suggest that Fyn facilitates mitotic spindle formation through the increase in microtubule polymerization, resulting in the acceleration of M-phase progression.

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Activation of Lyn Tyrosine Kinase through Decreased Membrane Cholesterol Levels during a Change in Its Membrane Distribution upon Cell Detachment

Cited by 10 publications

References 53 publications

Golgi Distribution of Lyn to Caveolin- and Giantin-Positive <i>cis</i>-Golgi Membranes and the Caveolin-Negative, TGN46-Positive <i>trans</i>-Golgi Network

Golgi Distribution of Lyn to Caveolin- and Giantin-Positive <i>cis</i>-Golgi Membranes and the Caveolin-Negative, TGN46-Positive <i>trans</i>-Golgi Network

Src Acts as an Effector for Ku70-dependent Suppression of Apoptosis through Phosphorylation of Ku70 at Tyr-530

Fyn Accelerates M Phase Progression by Promoting the Assembly of Mitotic Spindle Microtubules

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