Rap1 translates chemokine signals to integrin activation, cell polarization, and motility across vascular endothelium under flow

Shimonaka, Mika; Katagiri, Koko; Nakayama, Toshinori; Fujita, Naoya; Tsuruo, Takashi; Yoshie, Osamu; Kinashi, Tatsuo

doi:10.1083/jcb.200301133

Cited by 330 publications

(336 citation statements)

References 46 publications

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“…At the molecular level, we show that CRK proteins function to promote chemokine-dependent integrin activation by RAP1, a key event needed for adhesion to endothelial cells and subsequent diapedesis (27). Previous work has shown that CRK proteins signal through the RAP1 GEF C3G to regulate RAP1 activation (4,13,14,28).…”

Section: Discussionmentioning

confidence: 76%

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CRK proteins selectively regulate T cell migration into inflamed tissues

Huang¹,

Clarke²,

Karimi³

et al. 2015

J. Clin. Invest.

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R e s e a R c h a R t i c l e1 0 2 0 jci.org Volume 125 Number 3 March 2015 therapeutic implications, since they can carry out graftversus-leukemia (GVL) responses with minimal GVHD. Results Generation and characterization of T cell-specific CRK T cells (data not shown).Western blotting of purified CD4 + T cells from Dko and WT mice revealed that levels of CRKI, CRKII, and CRKL in the mutant T cells were reduced by at least 95% ( Figure 1A and data not shown), and flow cytometric analysis confirmed loss of CRK protein expression (Figure 1, B and C).CRK/CRKL Dko mice were born at Mendelian ratios, and spleen, thymus, and lymph nodes exhibited normal cellularity (data not shown). Thymocyte populations were similar in WT and Dko mice, indicating that T cell development proceeded normally ( Figure 1D). Peripheral lymphoid organs showed no differences in proportions of CD4 + and CD8 + T cells, naive (CD62L hi CD44 lo ), memory (CD62L lo CD44 hi ), or activated (CD69 hi ) T cell subsets (Figure 1, D and E, and data not shown). Thus, these mice represented a suitable source of mature CRK/CRKL Dko T cells for functional studies. CRK/CRKL-deficient T cells show defects in adhesion and polarization.Since CRK proteins control adhesion in non-hematopoietic cells (4), we first asked whether integrin-dependent adhesion is defective in CRK/CRKL Dko T cells. On plates coated with ICAM-1, the ligand for the β 2 integrin LFA-1, WT preactivated CD4 + T cells showed about 10% basal binding, which was increased 2-to 5-fold after stimulation with the chemokines CXCL12 or CCL21, or anti-CD3 (Figure 2A). CRK/CRKL Dko T cells showed a significant reduction in adhesion to ICAM-1 under both basal and stimulated conditions. Indeed, chemokine stimulation induced almost no increased adhesion in these cells. Treatment with PMA bypassed the defect. This is most likely because the defect in CRK/CRKL Dko cells lies upstream of PKC signaling in the pathway leading to integrin activation, though PKC activation could also drive a parallel

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Section: Discussionmentioning

confidence: 76%

“…We observed that in addition to playing a role in inside-out signaling pathways leading to integrin activation, CRK/CRKL Dko T cells polarized poorly in response to fibronectin ligation. This indicates that these proteins, like RAP1 (27,31), are also required for outside-in integrin signaling.…”

Section: Discussionmentioning

confidence: 98%

CRK proteins selectively regulate T cell migration into inflamed tissues

Huang¹,

Clarke²,

Karimi³

et al. 2015

J. Clin. Invest.

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“…The ras-like small GTPase Rap1 has recently emerged as an important regulator of early integrin activation by chemokine signals (42)(43)(44). Its crucial role in LFA-1 triggering is also evident from the latest description of a human genetic defect, leukocyte adhesion deficiency (LAD-III) (44), in which the expression of the Rap1 GEF, CalDAG-GEF-1, is impaired (45).…”

Section: Discussionmentioning

confidence: 99%

RhoA Is Involved in LFA-1 Extension Triggered by CXCL12 but Not in a Novel Outside-In LFA-1 Activation Facilitated by CXCL9

Pasvolsky

Grabovsky

Giagulli

et al. 2008

The Journal of Immunology

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Chemokines presented on endothelial tissues instantaneously trigger LFA-1-mediated arrest on ICAM-1 via rapid inside-out and outside-in (ligand-driven) LFA-1 activation. The GTPase RhoA was previously implicated in CCL21-triggered LFA-1 affinity triggering in murine T lymphocytes and in LFA-1-dependent adhesion strengthening to ICAM-1 on Peyer’s patch high endothelial venules stabilized over periods of at least 10 s. In this study, we show that a specific RhoA 23/40 effector region is vital for the initial LFA-1-dependent adhesions of lymphocytes on high endothelial venules lasting 1–3 s. Blocking the RhoA 23/40 region in human T lymphocytes in vitro also impaired the subsecond CXCL12-triggered LFA-1-mediated T cell arrest on ICAM-1 by eliminating the rapid induction of an extended LFA-1 conformational state. However, the inflammatory chemokine CXCL9 triggered robust LFA-1-mediated T lymphocyte adhesion to ICAM-1 at subsecond contacts independently of the RhoA 23/40 region. CXCL9 did not induce conformational changes in the LFA-1 ectodomain, suggesting that particular chemokines can activate LFA-1 through outside-in post ligand binding stabilization changes. Like CXCL9, the potent diacylglycerol-dependent protein kinase C agonist PMA was found to trigger LFA-1 adhesiveness to ICAM-1 also without inducing integrin extension or an a priori clustering and independently of the RhoA 23/40 region. Our results collectively suggest that the 23/40 region of RhoA regulates chemokine-induced inside-out LFA-1 extension before ligand binding, but is not required for a variety of chemokine and non-chemokine signals that rapidly strengthen LFA-1-ICAM-1 bonds without an a priori induction of high-affinity extended LFA-1 conformations.

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“…Rap1 is known to stimulate integrin activation, cytoskeleton rearrangements, and cell polarization both in the presence and absence of a chemokine gradient. 42,43 We observed that Rap1a and Rap1b depletion abolished the migratory advantage of MEC1-CD38H cells. This indicates that Rap1 activity contributes to the increased migration and adhesion linked to CD38 expression.…”

Section: Discussionmentioning

confidence: 76%

Anatomical Society Summer Meeting in Galway

2018

Journal of Anatomy

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Rap1 translates chemokine signals to integrin activation, cell polarization, and motility across vascular endothelium under flow

Cited by 330 publications

References 46 publications

CRK proteins selectively regulate T cell migration into inflamed tissues

CRK proteins selectively regulate T cell migration into inflamed tissues

RhoA Is Involved in LFA-1 Extension Triggered by CXCL12 but Not in a Novel Outside-In LFA-1 Activation Facilitated by CXCL9

Anatomical Society Summer Meeting in Galway

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