scholarly journals LIM and SH3 protein 1 localizes to the leading edge of protruding lamellipodia and regulates axon development

2020 ◽  
Vol 31 (24) ◽  
pp. 2718-2732
Author(s):  
Stephanie L. Pollitt ◽  
Kenneth R. Myers ◽  
Jin Yoo ◽  
James Q. Zheng
Keyword(s):  
Hippocampal Neurons ◽  
Binding Protein ◽  
Leading Edge ◽  
Actin Binding ◽  
Commissural Axon ◽  
Actin Binding Protein ◽  
Axon Elongation ◽  
Axon Development

This study reports that the actin-binding protein, LIM and SH3 Protein 1 (LASP1), regulates actin-based protrusions underlying axon elongation and branching in hippocampal neurons in culture. LASP1 also plays an important role in axon development in vivo, as loss of the Drosophila homologue LASP disrupts the commissural axon development.

scholarly journals Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin

2002 ◽  
Vol 159 (6) ◽  
pp. 993-1004 ◽  
Author(s):  
Christine L. Humphries ◽  
Heath I. Balcer ◽  
Jessica L. D'Agostino ◽  
Barbara Winsor ◽  
David G. Drubin ◽  
...  
Keyword(s):  
Binding Protein ◽  
Coiled Coil ◽  
Actin Binding ◽  
Complex Activity ◽  
Actin Binding Protein ◽  
Coiled Coil Domain ◽  
Allele Specific ◽  
And Function

Mechanisms for activating the actin-related protein 2/3 (Arp2/3) complex have been the focus of many recent studies. Here, we identify a novel mode of Arp2/3 complex regulation mediated by the highly conserved actin binding protein coronin. Yeast coronin (Crn1) physically associates with the Arp2/3 complex and inhibits WA- and Abp1-activated actin nucleation in vitro. The inhibition occurs specifically in the absence of preformed actin filaments, suggesting that Crn1 may restrict Arp2/3 complex activity to the sides of filaments. The inhibitory activity of Crn1 resides in its coiled coil domain. Localization of Crn1 to actin patches in vivo and association of Crn1 with the Arp2/3 complex also require its coiled coil domain. Genetic studies provide in vivo evidence for these interactions and activities. Overexpression of CRN1 causes growth arrest and redistribution of Arp2 and Crn1p into aberrant actin loops. These defects are suppressed by deletion of the Crn1 coiled coil domain and by arc35-26, an allele of the p35 subunit of the Arp2/3 complex. Further in vivo evidence that coronin regulates the Arp2/3 complex comes from the observation that crn1 and arp2 mutants display an allele-specific synthetic interaction. This work identifies a new form of regulation of the Arp2/3 complex and an important cellular function for coronin.

scholarly journals In vivo dynamics of the F-actin-binding protein neurabin-II

2000 ◽  
Vol 345 (2) ◽  
pp. 185 ◽  
Author(s):  
David J. STEPHENS ◽  
George BANTING
Keyword(s):  
Binding Protein ◽  
Actin Binding ◽  
Actin Binding Protein

Regulation of AMPA Receptor Recruitment by the Actin Binding Protein Drebrin in Cultured Hippocampal Neurons

2012 ◽  
Vol 1 (2) ◽  
pp. 153-160 ◽  
Author(s):  
Kenichi Kato ◽  
Tomoaki Shirao ◽  
Hiroyuki Yamazaki ◽  
Kazuyuki Imamura ◽  
Yuko Sekino
Keyword(s):  
Ampa Receptor ◽  
Hippocampal Neurons ◽  
Binding Protein ◽  
Actin Binding ◽  
Actin Binding Protein ◽  
Cultured Hippocampal Neurons

scholarly journals nPIST: A Novel Actin Binding Protein of trans-Golgi Network

2018 ◽  
Author(s):  
Swagata Das ◽  
Priyanka Dutta ◽  
Mohit Mazumder ◽  
Soma Seal ◽  
Kheerthana Duraivelan ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Purkinje Cells ◽  
Binding Protein ◽  
Vesicular Trafficking ◽  
Actin Binding ◽  
Perinuclear Region ◽  
Actin Binding Protein ◽  
Golgi Network

Abstractnpist is the neuronal isoform of PIST, a trans-golgi associated protein involved in major modulation of vesicular trafficking. nPIST interacts with glutamate delta2 receptor (GluRδ2) in Purkinje cells. Our study shows nPIST as a novel actin binding protein. Our structure based sequence analysis shows nPIST contains one WH2-like domain. Further our experimental analysis illustrates that fragment of nPIST consisting of WH2-like domain binds to actin. Moreover it was found that nPIST contains several regions involved in interaction with actin. The binding of nPIST to actin through multiple actin binding regions facilitated actin filament stabilization in vitro. In vivo, nPIST localized actin in perinuclear region as a blotch when ectopically expressed.

scholarly journals The Actin Binding Protein Plastin-3 Is Involved in the Pathogenesis of Acute Myeloid Leukemia

Cancers ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1663 ◽  
Author(s):  
Arne Velthaus ◽  
Kerstin Cornils ◽  
Jan K. Hennigs ◽  
Saskia Grüb ◽  
Hauke Stamm ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Binding Protein ◽  
Actin Binding ◽  
Bone Marrow Niche ◽  
Actin Binding Protein ◽  
Acute Myeloid

Leukemia-initiating cells reside within the bone marrow in specialized niches where they undergo complex interactions with their surrounding stromal cells. We have identified the actin-binding protein Plastin-3 (PLS3) as potential player within the leukemic bone marrow niche and investigated its functional role in acute myeloid leukemia. High expression of PLS3 was associated with a poor overall and event-free survival for AML patients. These findings were supported by functional in vitro and in vivo experiments. AML cells with a PLS3 knockdown showed significantly reduced colony numbers in vitro while the PLS3 overexpression variants resulted in significantly enhanced colony numbers compared to their respective controls. Furthermore, the survival of NSG mice transplanted with the PLS3 knockdown cells showed a significantly prolonged survival in comparison to mice transplanted with the control AML cells. Further studies should focus on the underlying leukemia-promoting mechanisms and investigate PLS3 as therapeutic target.

scholarly journals Microtubule-associated Protein 2c Reorganizes Both Microtubules and Microfilaments into Distinct Cytological Structures in an Actin-binding Protein-280–deficient Melanoma Cell Line

1997 ◽  
Vol 136 (4) ◽  
pp. 845-857 ◽  
Author(s):  
C. Casey Cunningham ◽  
Nicole Leclerc ◽  
Lisa A. Flanagan ◽  
Mei Lu ◽  
Paul A. Janmey ◽  
...  
Keyword(s):  
Melanoma Cell ◽  
Binding Protein ◽  
Growth Cones ◽  
Actin Binding ◽  
Actin Binding Protein ◽  
Low Concentrations ◽  
Rheologic Property ◽  
Neuronal Growth Cones

The emergence of processes from cells often involves interactions between microtubules and microfilaments. Interactions between these two cytoskeletal systems are particularly apparent in neuronal growth cones. The juvenile isoform of the neuronal microtubule-associated protein 2 (MAP2c) is present in growth cones, where we hypothesize it mediates interactions between microfilaments and microtubules. To approach this problem in vivo, we used the human melanoma cell, M2, which lacks actin-binding protein-280 (ABP-280) and forms membrane blebs, which are not seen in wild-type or ABP-transfected cells. The microinjection of tau or mature MAP2 rescued the blebbing phenotype; MAP2c not only caused cessation of blebbing but also induced the formation of two distinct cellular structures. These were actin-rich lamellae, which often included membrane ruffles, and microtubule-bearing processes. The lamellae collapsed after treatment with cytochalasin D, and the processes retracted after treatment with colchicine. MAP2c was immunocytochemically visualized in zones of the cell that were devoid of tubulin, such as regions within the lamellae and in association with membrane ruffles. In vitro rheometry confirmed that MAP2c is an efficient actin gelation protein capable of organizing actin filaments into an isotropic array at very low concentrations; tau and mature MAP2 do not share this rheologic property. These results suggest that MAP2c engages in functionally specific interactions not only with microtubules but also with microfilaments.

scholarly journals Mapping actin surfaces required for functional interactions in vivo.

1994 ◽  
Vol 126 (2) ◽  
pp. 423-432 ◽  
Author(s):  
D A Holtzman ◽  
K F Wertman ◽  
D G Drubin
Keyword(s):  
Binding Protein ◽  
Null Alleles ◽  
Actin Binding ◽  
Null Mutation ◽  
Wild Type ◽  
Actin Binding Protein ◽  
Functional Interactions ◽  
Type Gene ◽  
First Time

An in vivo strategy to identify amino acids of actin required for functional interactions with actin-binding proteins was developed. This approach is based on the assumption that an actin mutation that specifically impairs the interaction with an actin-binding protein will cause a phenotype similar to a null mutation in the gene that encodes the actin-binding protein. 21 actin mutations were analyzed in budding yeast, and specific regions of actin subdomain 1 were implicated in the interaction with fimbrin, an actin filament-bundling protein. Mutations in this actin subdomain were shown to be, like a null allele of the yeast fimbrin gene (SAC6), lethal in combination with null mutations in the ABP1 and SLA2 genes, and viable in combination with a null mutation in the SLA1 gene. Biochemical experiments with act1-120 actin (E99A, E100A) verified a defect in the fimbrin-actin interaction. Genetic interactions between mutant alleles of the yeast actin gene and null alleles of the SAC6, ABP1, SLA1, and SLA2 genes also demonstrated that the effects of the 21 actin mutations are diverse and allowed four out of seven pseudo-wild-type actin alleles to be distinguished from the wild-type gene for the first time, providing evidence for functional redundancy between different surfaces of actin.

scholarly journals The actin-binding protein Hip1R associates with clathrin during early stages of endocytosis and promotes clathrin assembly in vitro

2001 ◽  
Vol 154 (6) ◽  
pp. 1209-1224 ◽  
Author(s):  
Åsa E.Y. Engqvist-Goldstein ◽  
Robin A. Warren ◽  
Michael M. Kessels ◽  
James H. Keen ◽  
John Heuser ◽  
...  
Keyword(s):  
Binding Protein ◽  
Coiled Coil ◽  
Actin Binding ◽  
Live Cells ◽  
Cell Cortex ◽  
Coated Pits ◽  
Actin Binding Protein ◽  
Real Time Analysis

Huntingtin-interacting protein 1 related (Hip1R) is a novel component of clathrin-coated pits and vesicles and is a mammalian homologue of Sla2p, an actin-binding protein important for both actin organization and endocytosis in yeast. Here, we demonstrate that Hip1R binds via its putative central coiled-coil domain to clathrin, and provide evidence that Hip1R and clathrin are associated in vivo at sites of endocytosis. First, real-time analysis of Hip1R–YFP and DsRed–clathrin light chain (LC) in live cells revealed that these proteins show almost identical temporal and spatial regulation at the cell cortex. Second, at the ultrastructure level, immunogold labeling of ‘unroofed’ cells showed that Hip1R localizes to clathrin-coated pits. Third, overexpression of Hip1R affected the subcellular distribution of clathrin LC. Consistent with a functional role for Hip1R in endocytosis, we also demonstrated that it promotes clathrin cage assembly in vitro. Finally, we showed that Hip1R is a rod-shaped apparent dimer with globular heads at either end, and that it can assemble clathrin-coated vesicles and F-actin into higher order structures. In total, Hip1R's properties suggest an early endocytic function at the interface between clathrin, F-actin, and lipids.

scholarly journals Yeast actin-binding proteins: evidence for a role in morphogenesis.

1988 ◽  
Vol 107 (6) ◽  
pp. 2551-2561 ◽  
Author(s):  
D G Drubin ◽  
K G Miller ◽  
D Botstein
Keyword(s):  
Yeast Cell ◽  
Myosin Heavy Chain ◽  
Heavy Chain ◽  
Actin Filaments ◽  
Binding Proteins ◽  
Binding Protein ◽  
Actin Binding ◽  
Cortical Actin ◽  
Actin Binding Protein

Three yeast actin-binding proteins were identified using yeast actin filaments as an affinity matrix. One protein appears to be a yeast myosin heavy chain; it is dissociated from actin filaments by ATP, it is similar in size (200 kD) to other myosins, and antibodies directed against Dictyostelium myosin heavy chain bind to it. Immunofluorescence experiments show that a second actin-binding protein (67 kD) colocalizes in vivo with both cytoplasmic actin cables and cortical actin patches, the only identifiable actin structures in yeast. The cortical actin patches are concentrated at growing surfaces of the yeast cell where they might play a role in membrane and cell wall insertion, and the third actin-binding protein (85 kD) is only detected in association with these structures. This 85-kD protein is therefore a candidate for a determinant of growth sites. The in vivo role of this protein was tested by overproduction; this overproduction causes a reorganization of the actin cytoskeleton which in turn dramatically affects the budding pattern and spatial growth organization of the yeast cell.

scholarly journals The EGF receptor is an actin-binding protein.

1992 ◽  
Vol 119 (2) ◽  
pp. 349-355 ◽  
Author(s):  
J C den Hartigh ◽  
P M van Bergen en Henegouwen ◽  
A J Verkleij ◽  
J Boonstra
Keyword(s):  
Polyclonal Antibody ◽  
Synthetic Peptide ◽  
Egf Receptor ◽  
Binding Protein ◽  
Binding Domain ◽  
Actin Binding ◽  
Filamentous Actin ◽  
Actin Binding Protein ◽  
Actin Binding Domain

In a number of recent studies it has been shown that in vivo part of the EGF receptor (EGFR) population is associated to the actin filament system. In this paper we demonstrate that the purified EGFR can be cosedimented with purified filamentous actin (F-actin) indicating a direct association between EGFR and actin. A truncated EGFR, previously shown not to be associated to the cytoskeleton, was used as a control and this receptor did not cosediment with actin filaments. Determination of the actin-binding domain of the EGFR was done by measuring competition of either a polyclonal antibody or synthetic peptides on EGFR cosedimentation with F-actin. A synthetic peptide was made homologous to amino acid residues 984-996 (HL-33) of the EGFR which shows high homology with the actin-binding domain of Acanthamoeba profilin. A polyclonal antibody raised against HL-33 was found to prevent cosedimentation of EGFR with F-actin. This peptide HL-33 was shown to bind directly to actin in contrast with a synthetic peptide homologous to residues 1001-1013 (HL-34). During cosedimentation, HL-33 competed for actin binding of the EGFR and HL-34 did not, indicating that the EGFR contains one actin-binding site. These results demonstrate that the EGFR is an actin-binding protein which binds to actin via a domain containing amino acids residues 984-996.

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