Type Iγ phosphatidylinositol phosphate kinase targets and regulates focal adhesions

Ling, Kun; Doughman, Renee L.; Firestone, Ari J.; Bunce, Matthew W.; Anderson, Richard A.

doi:10.1038/nature01082

Cited by 414 publications

(556 citation statements)

References 31 publications

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“…Immunoblots were performed using standard conditions with polyclonal antibodies against PIPKI␥ (Ling et al, 2002) …”

Section: Protein Extraction Antibodies and Immunoblotsmentioning

confidence: 99%

“…Nonspecific binding was blocked with PBS containing 10% heat-inactivated fetal bovine serum and 0.25 mg/ml saponin at 4°C overnight. Cells were then stained for 30 min with ␣-PIPKI␥ (Ling et al, 2002) and costained with rhodamine-labeled phalloidin (Molecular Probes, Eugene, OR) and DAPI (Molecular Probes). FITC goat ␣-rabbit IgG (Jackson ImmunoResearch, West Grove, PA) was used as the secondary antibody.…”

Section: Immunofluorescencementioning

confidence: 99%

“…Wild-type GFP-PIPKI␥661 was generated by PCR amplification of a murine PIPKI␥661 cDNA (Ling et al, 2002; a generous gift from Dr. Richard Anderson) and subcloning into pEGFP (Clontech, Palo Alto, CA) as an N-terminal GFP fusion, which was then subcloned into the empty pMX retroviral vector (a generous gift from Dr. Clive Svendsen). Kinase-dead GFP-PIPKI␥661 (D253A; Ling et al, 2002) was generated by mutating wild-type pMX-GFP-PIPKI␥661 with the QuikChange site-directed mutagenesis kit (Stratagene, La Jolla, CA).…”

Section: Expression Constructsmentioning

confidence: 99%

“…Furthermore, PIPKI␥ mRNA is alternatively spliced to form PIPKI␥635 and PIPKI␥661, which differ by a 26-amino acid C-terminal extension. Previous work has shown that the PIPKI family of lipid kinases display distinct subcellular distributions in fibroblasts (Di Paolo et al, 2002;Ling et al, 2002;Doughman et al, 2003). PIPKI␣ targets to the leading edge of cells where it regulates membrane ruffling (Doughman et al, 2003), whereas PIPKI␥661 targets to focal adhesions and may regulate integrin-mediated adhesions during cell migration via an interaction with the cytoskeletal protein talin (Di Paolo et al, 2002;Ling et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…PIPKI␣ targets to the leading edge of cells where it regulates membrane ruffling (Doughman et al, 2003), whereas PIPKI␥661 targets to focal adhesions and may regulate integrin-mediated adhesions during cell migration via an interaction with the cytoskeletal protein talin (Di Paolo et al, 2002;Ling et al, 2002). However, PIPKI␥635 does not target to focal adhesions or interact with talin (Ling et al, 2002). Although PIPKI family members have been implicated in cell motility , the generation of localized PtdIns(4,5)P 2 and functions of PIPKI family members during neutrophil chemotaxis have not been explored.…”

Section: Introductionmentioning

confidence: 99%

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Type Iγ PIP Kinase Is a Novel Uropod Component that Regulates Rear Retraction during Neutrophil Chemotaxis

Lokuta

Senetar²,

Bennin

et al. 2007

MBoC

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Cell polarization is necessary for directed migration and leukocyte recruitment to inflamed tissues. Recent progress has been made in defining the molecular mechanisms that regulate chemoattractant-induced cell polarity during chemotaxis, including the contribution of phosphoinositide 3-kinase (PI3K)-dependent phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P 3 ] synthesis at the leading edge. However, less is known about the molecular composition of the cell rear and how the uropod functions during cell motility. Here, we demonstrate that phosphatidylinositol phosphate kinase type I␥ (PIPKI␥661), which generates PtdIns(4,5)P 2 , is enriched in the uropod during chemotaxis of primary neutrophils and differentiated HL-60 cells (dHL-60). Using time-lapse microscopy, we show that enrichment of PIPKI␥661 at the cell rear occurs early upon chemoattractant stimulation and is persistent during chemotaxis. Accordingly, we were able to detect enrichment of PtdIns(4,5)P 2 at the uropod during chemotaxis. Overexpression of kinase-dead PIPKI␥661 compromised uropod formation and rear retraction similar to inhibition of ROCK signaling, suggesting that PtdIns(4,5)P 2 synthesis is important to elicit the backness response during chemotaxis. Together, our findings identify a previously unknown function for PIPKI␥661 as a novel component of the backness signal that regulates rear retraction during chemotaxis.

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“…Immunoblots were performed using standard conditions with polyclonal antibodies against PIPKI␥ (Ling et al, 2002) …”

Section: Protein Extraction Antibodies and Immunoblotsmentioning

confidence: 99%

Section: Immunofluorescencementioning

confidence: 99%

Section: Expression Constructsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Type Iγ PIP Kinase Is a Novel Uropod Component that Regulates Rear Retraction during Neutrophil Chemotaxis

Lokuta

Senetar²,

Bennin

et al. 2007

MBoC

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Integrins: Signalling and Disease

Danen

2013

Encyclopedia of Life Sciences

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Cell‐surface adhesion molecules of the integrin family are not a passive glue, but rather are dynamic molecules that mediate the transfer of information across the membrane in both directions. Integrin‐mediated adhesion can be regulated in response to signals by clustering and conformational changes triggered at their cytoplasmic tails. Vice versa, integrins probe chemical and physical aspects of the extracellular environment and, in concert with growth factor receptors, activate signalling pathways in response. Integrin signalling controls cell survival, cell cycle progression, and differentiation, and the dynamic regulation of integrin‐mediated adhesion structures is critical for many forms of cell migration. Lastly, integrins contribute to the pathogenesis of a diverse array of acquired and hereditary diseases, and hence represent major targets for therapeutic intervention. Key Concepts: Integrins are heterodimeric transmembrane receptors that mediate cell adhesion to extracellular matrix proteins and other cells. Integrins cluster in cell–matrix or cell–cell adhesions where they assemble large multiprotein complexes including actin‐binding proteins. Integrin‐containing adhesions contain enzymes such as FAK, Src, Rac and many others as well as their upstream activators and downstream effectors allowing local activation of cellular signalling cascades. Through a series of conformational changes in mechanoresponsive proteins, integrin‐containing adhesions provide mechanical coupling between the environment and the cytoskeletal network. Signalling through various growth factor receptors is amplified in response to integrin‐mediated adhesion. The ability of integrins to modulate cellular shape and cytoarchitecture in accordance with environmental stiffness as well as their role in a variety of biochemical signal transduction cascades underlies their critical function in cell proliferation, survival, differentiation and motility. Defective integrin function underlies several human diseases. Integrins represent potential drug targets and therapeutic strategies based on interference with integrin‐mediated adhesion have entered clinical trials.

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Phosphoinositides – The Seven Species: Conversion and Cellular Roles

Lietha

2011

Encyclopedia of Life Sciences

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Phosphoinositides are phospholipids that contain a phosphorylated inositol head group. The position and number of phosphate groups varies, which results in seven phosphoinositide species. A large family of enzymes have evolved to specifically modify phosphoinositides. Phosphoinositide kinases and phosphatases modify the phosphorylation state of the inositol head group, whereas phospholipases hydrolyse phosphoinositides to release the soluble head group into the cytosol. The combined action of these enzymes produces the phosphoinositide signature of a cell, where certain membrane compartments are enriched or depleted of specific phosphoinositides. The cellular response of a certain phosphoinositide signature is mediated by phosphoinositide effectors. These effectors contain phosphoinositide recognition domains, which guide the effector to the appropriate location and in many cases also modulate their activity. Phosphoinositides play crucial roles in many cellular processes, including cell signalling, cytoskeletal rearrangements, vesicle transport and control of ion channels. Key Concepts: Phosphoinositides are phospholipids that contain a negatively charged phosphoinositol head group. Phosphoinositides are modified by phosphoinositide kinases, phosphatases and phospholipases. Specific phosphoinositides are enriched in different membrane compartments. Specific phosphoinositide recognition modules are linked to effectors. Tight regulation of generation and depletion of phosphoinositides allows spatio‐temporal control of complex cellular processes. PI3KI and PTEN, which control PtdIns(3,4,5)P 3 levels, belong to the most frequently mutated genes in cancer.

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Type Iγ phosphatidylinositol phosphate kinase targets and regulates focal adhesions

Cited by 414 publications

References 31 publications

Type Iγ PIP Kinase Is a Novel Uropod Component that Regulates Rear Retraction during Neutrophil Chemotaxis

Type Iγ PIP Kinase Is a Novel Uropod Component that Regulates Rear Retraction during Neutrophil Chemotaxis

Integrins: Signalling and Disease

Phosphoinositides – The Seven Species: Conversion and Cellular Roles

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