VAMP4‐ and VAMP7‐expressing vesicles are both required for cytotoxic granule exocytosis in NK cells

Krzewski, Konrad; Gil-Krzewska, Aleksandra; Watts, James J.; Stern, Joel N. H.; Strominger, Jack L.

doi:10.1002/eji.201141582

Cited by 33 publications

(41 citation statements)

References 27 publications

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“…Consistently, FLAGRab40b colocalizes with VAMP4-secretory vesicles at the periphery of the cell (Fig. 5G-L, arrows), suggesting that Rab40b organelles contain VAMP4 (Kakhlon et al, 2006;Krzewski et al, 2011;Steegmaier et al, 1999). Similarly, VAMP4 also colocalized with MMP2-Myc and MMP9-Myc (supplementary material Fig.…”

Section: Rab40b Gtpase Is Required For Mmp2 and Mmp9 Secretionsupporting

confidence: 63%

Rab40b regulates MMP2 and MMP9 trafficking during invadopodia formation and breast cancer cell invasion

Jacob

Jing

Lee³

et al. 2013

Journal of Cell Science

125

136

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SummaryInvadopodia-dependent degradation of the basement membrane plays a major role during metastasis of breast cancer cells. Basement membrane degradation is mediated by targeted secretion of various matrix metalloproteinases (MMPs). Specifically, MMP2 and MMP9 (MMP2/9) possess the ability to hydrolyze components of the basement membrane and regulate various aspects of tumor growth and metastasis. However, the membrane transport machinery that mediates targeting of MMP2/9 to the invadopodia during cancer cell invasion remains to be defined. Because Rab GTPases are key regulators of membrane transport, we screened a human Rab siRNA library and identified Rab40b GTPase as a protein required for secretion of MMP2/9. We also have shown that Rab40b functions during at least two distinct steps of MMP2/9 transport. Here, we demonstrate that Rab40b is required for MMP2/9 sorting into VAMP4-containing secretory vesicles. We also show that Rab40b regulates transport of MMP2/9 secretory vesicles during invadopodia formation and is required for invadopodia-dependent extracellular matrix degradation. Finally, we demonstrate that Rab40b is also required for breast cancer cell invasion in vitro. On the basis of these findings, we propose that Rab40b mediates trafficking of MMP2/9 during invadopodia formation and metastasis of breast cancer cells.

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Section: Rab40b Gtpase Is Required For Mmp2 and Mmp9 Secretionsupporting

confidence: 63%

Rab40b regulates MMP2 and MMP9 trafficking during invadopodia formation and breast cancer cell invasion

Jacob

Jing

Lee³

et al. 2013

Journal of Cell Science

125

136

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“…Both nontargeting shRNA and scrambled siRNA are collectively referred to as control (CTRL) RNA interference (RNAi). Generation of lentivirus particles and infection of YTS cells was done as described by Krzewski et al 16 siRNA was delivered to ex vivo isolated NK cells by nucleofection using Nucleofector II (Lonza), and the cells were analyzed 72 hours after the procedure.…”

Section: Rnai Constructsmentioning

confidence: 99%

“…[8][9][10][11] Knowledge about the proteins involved in regulation of lytic granule release is very limited. Soluble N-ethylmaleimidesensitive factor attachment protein receptor proteins and small guanosine triphosphatases have been shown to be required for NK-cell degranulation and tumor killing, [12][13][14][15][16] but the granule-specific protein machinery involved in NK-cell exocytosis is poorly understood.…”

Section: Introductionmentioning

confidence: 99%

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LAMP1/CD107a is required for efficient perforin delivery to lytic granules and NK-cell cytotoxicity

Krzewski

Gil-Krzewska

Nguyen

et al. 2013

Blood

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125

105

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Key Points• LAMP1 silencing inhibits cytotoxicity of human NK cells.• LAMP1 is important for perforin trafficking to the lytic granules and granule movement.Secretory lysosomes of natural killer (NK) cells, containing perforin and granzymes, are indispensable for NK-cell cytotoxicity because their release results in the induction of target-cell apoptosis. Lysosome-associated membrane protein (LAMP) 1/CD107a is used as a marker for NK-cell degranulation, but its role in NK-cell biology is unknown. We show that LAMP1 silencing causes inhibition of NK-cell cytotoxicity, as LAMP1 RNA interference (RNAi) cells fail to deliver granzyme B to target cells. Reduction of LAMP1 expression affects the movement of lytic granules and results in decreased levels of perforin, but not granzyme B, in the granules. In LAMP1 RNAi cells, more perforin is retained outside of lysosomal compartments in trans-Golgi network-derived transport vesicles. Disruption of expression of LAMP1 binding partner, adaptor protein 1 (AP-1) sorting complex, also causes retention of perforin in the transport vesicles and inhibits cytotoxicity, indicating that the interaction between AP-1 sorting complex and LAMP1 on the surface of the transport vesicles is important for perforin trafficking to lytic granules. We conclude that the decreased level of perforin in lytic granules of LAMP1-deficient cells, combined with disturbed motility of the lytic granules, leads to the inability to deliver apoptosis-inducing granzyme B to target cells and to inhibition of NK-cell cytotoxicity. (Blood. 2013;121(23):4672-4683)

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“…Although a number of SNARE proteins have been shown to influence individual LG maturation steps [8][9][10] the molecular composition of the SNARE complex mediating the most important step of LG function, the final fusion with the plasma membrane to deliver the lethal hit that enables target cell killing, remains unknown. For the t-SNAREs several candidates have been proposed 7,11 , whereas no candidate v-SNARE protein has emerged so far.…”

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confidence: 99%

Synaptobrevin2 is the v-SNARE required for cytotoxic T-lymphocyte lytic granule fusion

et al. 2013

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Cytotoxic T lymphocytes kill virus-infected and tumorigenic target cells through the release of perforin and granzymes via fusion of lytic granules at the contact site, the immunological synapse. It has been postulated that this fusion process is mediated by non-neuronal members of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex protein family. Here, using a synaptobrevin2-monomeric red fluorescence protein knock-in mouse we demonstrate that, surprisingly, the major neuronal v-SNARE synaptobrevin2 is expressed in cytotoxic T lymphocytes and exclusively localized on granzyme B-containing lytic granules. Cleavage of synaptobrevin2 by tetanus toxin or ablation of the synaptobrevin2 gene leads to a complete block of lytic granule exocytosis while leaving upstream events unaffected, identifying synaptobrevin2 as the v-SNARE responsible for the fusion of lytic granules at the immunological synapse.

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VAMP4‐ and VAMP7‐expressing vesicles are both required for cytotoxic granule exocytosis in NK cells

Cited by 33 publications

References 27 publications

Rab40b regulates MMP2 and MMP9 trafficking during invadopodia formation and breast cancer cell invasion

Rab40b regulates MMP2 and MMP9 trafficking during invadopodia formation and breast cancer cell invasion

LAMP1/CD107a is required for efficient perforin delivery to lytic granules and NK-cell cytotoxicity

Synaptobrevin2 is the v-SNARE required for cytotoxic T-lymphocyte lytic granule fusion

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