An actin-based wave generator organizes cell motility (2007)
Weiner, Orion D, Marganski, William A, Wu, Lani F, Altschuler, Steven J, Kirschner, Marc W
Although many of the regulators of actin assembly are known, we do not understand how these components act together to organize cell shape and movement. To address this question, we analyzed the...
An Actin-Based Wave Generator Organizes Cell Motility (2007)
Orion D. Weiner, William A. Marganski, Lani F. Wu, Steven J. Altschuler, Marc W. Kirschner
Although many of the regulators of actin assembly are known, we do not understand how these components act together to organize cell shape and movement. To address this question, we analyzed the...
Orion D. Weiner, Maike C. Rentel, Alex Ott, Glenn E. Brown, Mark Jedrychowski, Michael B. Yaffe, ...
The authors describe a group of multiprotein complexes, scaffolded by Hem-1 (a subset of which also contain WAVE2), that are involved in generating and maintaining cellular polarity in neutrophils.
Orion D. Weiner, Maike C. Rentel, Alex Ott, Glenn E. Brown, Mark Jedrychowski, Michael B. Yaffe, ...
Migrating cells need to make different actin assemblies at the cell's leading and trailing edges and to maintain physical separation of signals for these assemblies. This asymmetric control of...
Campellone, Kenneth Geno, Rankin, Susannah, Pawson, Tony, Kirschner, Marc W., Tipper, Donald J., Leong, John M.
Enteropathogenic Escherichia coli (EPEC) translocates effector proteins into mammalian cells to promote reorganization of the cytoskeleton into filamentous actin pedestals. One effector, Tir, is a...
Ethan Lee, Adrian Salic, Roland Krüger, Reinhart Heinrich, Marc W. Kirschner
Wnt signaling is important in both oncogenesis and development. Mathematical modeling predicts several unusual features of the Wnt pathway, some of which are tested experimentally.
Ethan Lee, Adrian Salic, Roland Krüger, Reinhart Heinrich, Marc W. Kirschner
Wnt signaling plays an important role in both oncogenesis and development. Activation of the Wnt pathway results in stabilization of the transcriptional coactivator β-catenin. Recent studies have...
Ethan Lee, Adrian Salic, Reinhart Heinrich, Marc W. Kirschner
Wnt signaling plays an important role in both oncogenesis and development. Activation of the Wnt pathway results in stabilization of the transcriptional coactivator b-catenin. Recent studies have...
Mitotic inactivation of a human SWI/SNF chromatin remodeling complex (1998)
Sif, Saïd, Stukenberg, P. Todd, Kirschner, Marc W., Kingston, Robert E.
Tubulin requires tau for growth onto microtubule initiating sites (1976)
Witman, George B., Cleveland, Don W., Weingarten, Murray D., Kirschner, Marc W.
Tubulin purified by phosphocellulose chromatography and free of accessory proteins will not form microtubules in the absence or presence of microtubule initiating sites (flagellar microtubules). The...
Yamaguchi, Hideki, Miki, Hiroaki, Suetsugu, Shiro, Ma, Le, Kirschner, Marc W., Takenawa, Tadaomi
All WASP family proteins share a common C terminus that consists of the verprolin homology domain (V), cofilin homology domain (C), and acidic region (A), through which they activate Arp2/3...
Mead, Paul E., Zhou, Yi, Lustig, Kevin D., Huber, Tara L., Kirschner, Marc W., Zon, Leonard I.
We have developed a technique, fast retrieval of gel shift activities (FROGS), that allows for the rapid isolation of proteins that interact with DNA. Using this technique, we have isolated two...
Zhou, Bin-Bing, Li, Honglin, Yuan, Junying, Kirschner, Marc W.
The activation of cyclin-dependent kinases (cdks) has been implicated in apoptosis induced by various stimuli. We find that the Fas-induced activation of cdc2 and cdk2 in Jurkat cells is not...
The Arp2/3 complex mediates actin polymerization induced by the small GTP-binding protein Cdc42
Ma, Le, Rohatgi, Rajat, Kirschner, Marc W.
The small GTP-binding protein Cdc42 is thought to induce filopodium formation by regulating actin polymerization at the cell cortex. Although several Cdc42-binding proteins have been identified and...
A chemical inhibitor of N-WASP reveals a new mechanism for targeting protein interactions
Peterson, Jeffrey R., Lokey, R. Scott, Mitchison, Timothy J., Kirschner, Marc W.
Cell morphology and motility are governed largely by complex signaling networks that ultimately engage the actin cytoskeleton. Understanding how individual circuits contribute to the process of...
Ho, Hsin-Yi Henry, Rohatgi, Rajat, Ma, Le, Kirschner, Marc W.
The Neuronal Wiskott–Aldrich syndrome protein (N-WASP) has emerged as a central regulator of the actin cytoskeleton with abilities to integrate multiple upstream signal inputs and transmit them to...
CDC20/CDH1 activates the anaphase-promoting complex (APC) and targets various substrates for degradation, thereby allowing the ordered progression through mitosis and G1. We have found multiple...
Chabes, Anna Lena, Pfleger, Cathie M., Kirschner, Marc W., Thelander, Lars
Ribonucleotide reductase consists of two nonidentical proteins, R1 and R2, and catalyzes the rate-limiting step in DNA precursor synthesis: the reduction of ribonucleotides to deoxyribonucleotides. A...
WAVE2 deficiency reveals distinct roles in embryogenesis and Rac-mediated actin-based motility
Yan, Catherine, Martinez-Quiles, Narcisa, Eden, Sharon, Shibata, Tomoyuki, Takeshima, Fuminao, Shinkura, Reiko, ...
The Wiskott–Aldrich syndrome related protein WAVE2 is implicated in the regulation of actin-cytoskeletal reorganization downstream of the small Rho GTPase, Rac. We inactivated the WAVE2 gene by...
Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS
Gerber, Scott A., Rush, John, Stemman, Olaf, Kirschner, Marc W., Gygi, Steven P.
A need exists for technologies that permit the direct quantification of differences in protein and posttranslationally modified protein expression levels. Here we present a strategy for the absolute...
The Roles of APC and Axin Derived from Experimental and Theoretical Analysis of the Wnt Pathway
Lee, Ethan, Salic, Adrian, Krüger, Roland, Heinrich, Reinhart, Kirschner, Marc W
Wnt signaling plays an important role in both oncogenesis and development. Activation of the Wnt pathway results in stabilization of the transcriptional coactivator β-catenin. Recent studies have...
The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1
Pfleger, Cathie M., Kirschner, Marc W.
The ordered progression through the cell cycle depends on regulating the abundance of several proteins through ubiquitin-mediated proteolysis. Degradation is precisely timed and specific. One key...
Shen, Minhui, Stukenberg, P. Todd, Kirschner, Marc W., Lu, Kun Ping
Phosphorylation of mitotic proteins on the Ser/Thr-Pro motifs has been shown to play an important role in regulating mitotic progression. Pin1 is a novel essential peptidyl–prolyl isomerase...
Fang, Guowei, Yu, Hongtao, Kirschner, Marc W.
The spindle assembly checkpoint mechanism delays anaphase initiation until all chromosomes are aligned at the metaphase plate. Activation of the anaphase-promoting complex (APC) by binding of CDC20...
Mitotic inactivation of a human SWI/SNF chromatin remodeling complex
Sif, Saïd, Stukenberg, P. Todd, Kirschner, Marc W., Kingston, Robert E.
During mitosis, chromatin is condensed into mitotic chromosomes and transcription is inhibited, processes that might be opposed by the chromatin remodeling activity of the SWI/SNF complexes. Brg1 and...
Purification and architecture of the ubiquitous Wave complex
Gautreau, Alexis, Ho, Hsin-yi H., Li, Jiaxu, Steen, Hanno, Gygi, Steven P., Kirschner, Marc W.
The Wave proteins are major activators of the Arp2/3 complex. The ubiquitous Wave-2 is required for actin polymerization at the leading edge of migrating cells. Here we purify Wave-2 from HeLa cells....
Kirschner, Marc W., Williams, Robley C., Weingarten, Murray, Gerhart, John C.
Depolymerization products of purified microtubules from porcine brain were examined by sedimentation analysis and electron microscopy. The complete depolymerization mixture exhibited 36S and 6S...
Martinez-Quiles, Narcisa, Ho, Hsin-Yi Henry, Kirschner, Marc W., Ramesh, Narayanaswamy, Geha, Raif S.
The Arp2/3 complex can be independently activated to initiate actin polymerization by the VCA domain of WASP family members and by the acidic N-terminal and F-actin-binding repeat region of...
Margalit, Danielle N., Romberg, Laura, Mets, Rebecca B., Hebert, Alan M., Mitchison, Timothy J., Kirschner, Marc W., ...
FtsZ, the ancestral homolog of eukaryotic tubulins, is a GTPase that assembles into a cytokinetic ring structure essential for cell division in prokaryotic cells. Similar to tubulin, purified FtsZ...
Weiner, Orion D, Rentel, Maike C, Ott, Alex, Brown, Glenn E, Jedrychowski, Mark, Yaffe, Michael B, ...
Migrating cells need to make different actin assemblies at the cell's leading and trailing edges and to maintain physical separation of signals for these assemblies. This asymmetric control of...
Inhibition of Cdh1–APC by the MAD2-related protein MAD2L2: a novel mechanism for regulating Cdh1
Pfleger, Cathie M., Salic, Adrian, Lee, Ethan, Kirschner, Marc W.
Exit from mitosis requires the degradation of regulatory proteins including the mitotic cyclins and securin through ubiquitination by the anaphase promoting complex (APC) bound to Cdc20 or Cdh1....
Substrate recognition by the Cdc20 and Cdh1 components of the anaphase-promoting complex
Pfleger, Cathie M., Lee, Ethan, Kirschner, Marc W.
The specificity of ubiquitin-mediated protein degradation with regards to the selection of substrates to be polyubiquitinated has only been determined rather recently. Substrate targeting by the...
Yamaguchi, Hideki, Miki, Hiroaki, Suetsugu, Shiro, Ma, Le, Kirschner, Marc W., Takenawa, Tadaomi
All WASP family proteins share a common C terminus that consists of the verprolin homology domain (V), cofilin homology domain (C), and acidic region (A), through which they activate Arp2/3...
Mead, Paul E., Zhou, Yi, Lustig, Kevin D., Huber, Tara L., Kirschner, Marc W., Zon, Leonard I.
We have developed a technique, fast retrieval of gel shift activities (FROGS), that allows for the rapid isolation of proteins that interact with DNA. Using this technique, we have isolated two...
Zhou, Bin-Bing, Li, Honglin, Yuan, Junying, Kirschner, Marc W.
The activation of cyclin-dependent kinases (cdks) has been implicated in apoptosis induced by various stimuli. We find that the Fas-induced activation of cdc2 and cdk2 in Jurkat cells is not...
The Arp2/3 complex mediates actin polymerization induced by the small GTP-binding protein Cdc42
Ma, Le, Rohatgi, Rajat, Kirschner, Marc W.
The small GTP-binding protein Cdc42 is thought to induce filopodium formation by regulating actin polymerization at the cell cortex. Although several Cdc42-binding proteins have been identified and...
A chemical inhibitor of N-WASP reveals a new mechanism for targeting protein interactions
Peterson, Jeffrey R., Lokey, R. Scott, Mitchison, Timothy J., Kirschner, Marc W.
Cell morphology and motility are governed largely by complex signaling networks that ultimately engage the actin cytoskeleton. Understanding how individual circuits contribute to the process of...
Ho, Hsin-Yi Henry, Rohatgi, Rajat, Ma, Le, Kirschner, Marc W.
The Neuronal Wiskott–Aldrich syndrome protein (N-WASP) has emerged as a central regulator of the actin cytoskeleton with abilities to integrate multiple upstream signal inputs and transmit them to...
CDC20/CDH1 activates the anaphase-promoting complex (APC) and targets various substrates for degradation, thereby allowing the ordered progression through mitosis and G1. We have found multiple...
Chabes, Anna Lena, Pfleger, Cathie M., Kirschner, Marc W., Thelander, Lars
Ribonucleotide reductase consists of two nonidentical proteins, R1 and R2, and catalyzes the rate-limiting step in DNA precursor synthesis: the reduction of ribonucleotides to deoxyribonucleotides. A...
WAVE2 deficiency reveals distinct roles in embryogenesis and Rac-mediated actin-based motility
Yan, Catherine, Martinez-Quiles, Narcisa, Eden, Sharon, Shibata, Tomoyuki, Takeshima, Fuminao, Shinkura, Reiko, ...
The Wiskott–Aldrich syndrome related protein WAVE2 is implicated in the regulation of actin-cytoskeletal reorganization downstream of the small Rho GTPase, Rac. We inactivated the WAVE2 gene by...
Absolute quantification of proteins and phosphoproteins from cell lysates by tandem MS
Gerber, Scott A., Rush, John, Stemman, Olaf, Kirschner, Marc W., Gygi, Steven P.
A need exists for technologies that permit the direct quantification of differences in protein and posttranslationally modified protein expression levels. Here we present a strategy for the absolute...
The Roles of APC and Axin Derived from Experimental and Theoretical Analysis of the Wnt Pathway
Lee, Ethan, Salic, Adrian, Krüger, Roland, Heinrich, Reinhart, Kirschner, Marc W
Wnt signaling plays an important role in both oncogenesis and development. Activation of the Wnt pathway results in stabilization of the transcriptional coactivator β-catenin. Recent studies have...
Inhibition of Cdh1–APC by the MAD2-related protein MAD2L2: a novel mechanism for regulating Cdh1
Pfleger, Cathie M., Salic, Adrian, Lee, Ethan, Kirschner, Marc W.
Exit from mitosis requires the degradation of regulatory proteins including the mitotic cyclins and securin through ubiquitination by the anaphase promoting complex (APC) bound to Cdc20 or Cdh1....
Substrate recognition by the Cdc20 and Cdh1 components of the anaphase-promoting complex
Pfleger, Cathie M., Lee, Ethan, Kirschner, Marc W.
The specificity of ubiquitin-mediated protein degradation with regards to the selection of substrates to be polyubiquitinated has only been determined rather recently. Substrate targeting by the...
The KEN box: an APC recognition signal distinct from the D box targeted by Cdh1
Pfleger, Cathie M., Kirschner, Marc W.
The ordered progression through the cell cycle depends on regulating the abundance of several proteins through ubiquitin-mediated proteolysis. Degradation is precisely timed and specific. One key...
Shen, Minhui, Stukenberg, P. Todd, Kirschner, Marc W., Lu, Kun Ping
Phosphorylation of mitotic proteins on the Ser/Thr-Pro motifs has been shown to play an important role in regulating mitotic progression. Pin1 is a novel essential peptidyl–prolyl isomerase...
Fang, Guowei, Yu, Hongtao, Kirschner, Marc W.
The spindle assembly checkpoint mechanism delays anaphase initiation until all chromosomes are aligned at the metaphase plate. Activation of the anaphase-promoting complex (APC) by binding of CDC20...
Mitotic inactivation of a human SWI/SNF chromatin remodeling complex
Sif, Saïd, Stukenberg, P. Todd, Kirschner, Marc W., Kingston, Robert E.
During mitosis, chromatin is condensed into mitotic chromosomes and transcription is inhibited, processes that might be opposed by the chromatin remodeling activity of the SWI/SNF complexes. Brg1 and...
Purification and architecture of the ubiquitous Wave complex
Gautreau, Alexis, Ho, Hsin-yi H., Li, Jiaxu, Steen, Hanno, Gygi, Steven P., Kirschner, Marc W.
The Wave proteins are major activators of the Arp2/3 complex. The ubiquitous Wave-2 is required for actin polymerization at the leading edge of migrating cells. Here we purify Wave-2 from HeLa cells....
Kirschner, Marc W., Williams, Robley C., Weingarten, Murray, Gerhart, John C.
Depolymerization products of purified microtubules from porcine brain were examined by sedimentation analysis and electron microscopy. The complete depolymerization mixture exhibited 36S and 6S...
Martinez-Quiles, Narcisa, Ho, Hsin-Yi Henry, Kirschner, Marc W., Ramesh, Narayanaswamy, Geha, Raif S.
The Arp2/3 complex can be independently activated to initiate actin polymerization by the VCA domain of WASP family members and by the acidic N-terminal and F-actin-binding repeat region of...
Margalit, Danielle N., Romberg, Laura, Mets, Rebecca B., Hebert, Alan M., Mitchison, Timothy J., Kirschner, Marc W., ...
FtsZ, the ancestral homolog of eukaryotic tubulins, is a GTPase that assembles into a cytokinetic ring structure essential for cell division in prokaryotic cells. Similar to tubulin, purified FtsZ...
Steen, Hanno, Jebanathirajah, Judith A., Springer, Michael, Kirschner, Marc W.
Qualitative and quantitative information are crucial to a detailed understanding of the function of protein phosphorylation. MS is now becoming a quantitative approach to analyze protein...
Weiner, Orion D, Rentel, Maike C, Ott, Alex, Brown, Glenn E, Jedrychowski, Mark, Yaffe, Michael B, ...
Migrating cells need to make different actin assemblies at the cell's leading and trailing edges and to maintain physical separation of signals for these assemblies. This asymmetric control of...
An Actin-Based Wave Generator Organizes Cell Motility
Weiner, Orion D, Marganski, William A, Wu, Lani F, Altschuler, Steven J, Kirschner, Marc W
Although many of the regulators of actin assembly are known, we do not understand how these components act together to organize cell shape and movement. To address this question, we analyzed the...
Campellone, Kenneth G., Rankin, Susannah, Pawson, Tony, Kirschner, Marc W., Tipper, Donald J., Leong, John M.
Enteropathogenic Escherichia coli (EPEC) translocates effector proteins into mammalian cells to promote reorganization of the cytoskeleton into filamentous actin pedestals. One effector, Tir, is a...
The relationship between evolutionary and physiological variation in hemoglobin
Milo, Ron, Hou, Jennifer H., Springer, Michael, Brenner, Michael P., Kirschner, Marc W.
Physiological and evolutionary adaptations operate at very different time scales. Nevertheless, there are reasons to believe there should be a strong relationship between the two, as together they...
Le Ma,, Cantley, Lewis C., Janmey, Paul A., Kirschner, Marc W.
Both phosphoinositides and small GTP-binding proteins of the Rho family have been postulated to regulate actin assembly in cells. We have reconstituted actin assembly in response to these signals in...
Mechanism of N-Wasp Activation by Cdc42 and Phosphatidylinositol 4,5-Bisphosphate
Rohatgi, Rajat, Ho, Hsin-yi Henry, Kirschner, Marc W.
Neuronal Wiskott-Aldrich Syndrome protein (N-WASP) transmits signals from Cdc42 to the nucleation of actin filaments by Arp2/3 complex. Although full-length N-WASP is a weak activator of Arp2/3...
Physiological regulation of β-catenin stability by Tcf3 and CK1ε
Lee, Ethan, Salic, Adrian, Kirschner, Marc W.
The wnt pathway regulates the steady state level of β-catenin, a transcriptional coactivator for the Tcf3/Lef1 family of DNA binding proteins. We demonstrate that Tcf3 can inhibit β-catenin...
Steen, Judith A. J., Steen, Hanno, Georgi, Ann, Parker, Kenneth, Springer, Michael, Kirchner, Marc, ...
The anaphase promoting complex (APC) controls the degradation of proteins during exit from mitosis and entry into S-phase. The activity of the APC is regulated by phosphorylation during mitosis....
Large-scale detection of ubiquitination substrates using cell extracts and protein microarrays
Merbl, Yifat, Kirschner, Marc W.
Identification of protein targets of post-translational modification is an important analytical problem in biology. Protein microarrays exposed to cellular extracts could offer a rapid and convenient...