Related Experiment Video

Updated: Jul 2, 2025

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration
10:43

Methods to Study Mrp4-containing Macromolecular Complexes in the Regulation of Fibroblast Migration

Published on: May 19, 2016

6.8K

Regulation of microtubule nucleation in mouse bone marrow-derived mast cells by ARF GTPase-activating protein GIT2

Vadym Sulimenko1, Vladimíra Sládková1, Tetyana Sulimenko1

  • 1Laboratory of Biology of Cytoskeleton, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, Czechia.

Frontiers in Immunology
|February 19, 2024
PubMed

Insights

G protein-coupled receptor kinase-interacting protein 2 (GIT2) regulates microtubule nucleation in mast cells. This protein acts as a negative regulator, impacting degranulation and cell movement during allergic responses.

Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • Mast cell activation via FcϵRI aggregation initiates signaling cascades, including calcium influx and mediator release.
  • Microtubules are crucial for mast cell degranulation, but their remodeling regulation is not fully understood.

Purpose of the Study:

  • To investigate the role of G protein-coupled receptor kinase-interacting protein 2 (GIT2) in regulating microtubule dynamics during mast cell activation.
  • To elucidate the mechanisms by which GIT2 influences centrosomal microtubule nucleation and mast cell function.

Main Methods:

  • FcϵRI-mediated activation of bone marrow-derived mast cells.
  • Immunofluorescence microscopy to assess microtubule nucleation and GIT2 localization.
  • GIT2 knockdown using siRNA and rescue experiments.
  • Analysis of mast cell degranulation and chemotaxis.

Main Results:

  • FcϵRI aggregation enhances centrosomal microtubule nucleation in mast cells.
  • GIT2 associates with centrosomes and γ-tubulin complex proteins.
  • GIT2 depletion increases microtubule nucleation, indicating a negative regulatory role.
  • Phosphorylation of GIT2 by protein kinase C promotes microtubule nucleation during activation.
  • GIT2 modulates antigen-induced degranulation and chemotaxis.

Conclusions:

  • GIT2 is a novel negative regulator of centrosomal microtubule nucleation in activated mast cells.
  • GIT2's phosphorylation status influences its centrosomal localization and function.
  • GIT2 plays a significant role in mast cell degranulation and migration processes.
Keywords:
G protein-coupled receptor kinase-interacting protein 2 (GIT2)centrosomemast cellsmicrotubule nucleationprotein kinase C (PKC)

More Related Videos

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells10:25

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells

Published on: August 13, 2016

11.1K
High-resolution Imaging and Analysis of Individual Astral Microtubule Dynamics in Budding Yeast10:23

High-resolution Imaging and Analysis of Individual Astral Microtubule Dynamics in Budding Yeast

Published on: April 20, 2017

9.5K

Related Concept Videos

Generation of Straight or Branched Actin Filaments01:14

Generation of Straight or Branched Actin Filaments

The straight or branched structure formation of actin filaments is controlled by nucleating proteins such as the formins and Arp2/3 complex. Formin-mediated assembly results in straight filaments, whereas Arp2/3 protein complex-mediated assembly results in branched actin filaments.
Arp2/3 Complex
Arp2/3 complex is a seven-subunit complex consisting of two proteins similar to actin- Arp2 and Arp3, and five other subunits that help keep Arp2 and Arp3 inactive. When required, the complex is...
2.9K
Microtubule Formation01:23

Microtubule Formation

Microtubules are dynamic structures that undergo continuous assembly and disassembly. They originate from specialized multi-protein complexes known as microtubule organizing centers or MTOCs. Within the MTOC, the point of origin of the microtubule is known as the minus end, while the end radiating outward is the plus end. Microtubules serve two primary functions — the organization of spindle complexes to separate sister chromatids during mitotic or meiotic cell division and the formation...
5.7K
Microtubule Instability02:17

Microtubule Instability

Microtubules are hollow cylindrical filaments having a diameter of approximately 25 nm and a length that varies from 200 nm to 25 μm. GTP-bound tubulin subunits form αβ-heterodimers for microtubule assembly. These core building blocks interact longitudinally, polymerizing into protofilaments. The protofilaments then interact with one another through lateral bonding forces to form stable cylindrical microtubules. These cylindrical filaments are dynamic as they undergo repeated...
5.1K
Mechanism of Lamellipodia Formation01:31

Mechanism of Lamellipodia Formation

Cells migrating in response to external stimuli form lamellipodia, which are thin membrane protrusions supported by a mesh of linked, branched, or unbranched actin filaments. These actin filaments interact with myosin motor proteins, creating the dynamic actomyosin complex within the cytoskeleton. Contractility, or the ability to generate contractile stress, is inherent to the actomyosin complex. It helps cells detect the stiffness of the surrounding ECM and exert contractile force for...
2.6K
Microtubule Associated Proteins (MAPs)01:42

Microtubule Associated Proteins (MAPs)

Microtubule function and architecture are regulated by an array of specialized proteins called microtubule-associated proteins or MAPs. These proteins are widespread across different organisms and have conserved protein motifs, like the multi-TOG domain for tubulin binding found in the CLASP family of MAPs. Some MAPs are lineage-specific based on their conserved domains. Their functions depend upon the cytoskeletal architecture and cell type they are located within. In-plant cells, a specific...
4.3K
Destabilization of Microtubules01:45

Destabilization of Microtubules

The destabilization of microtubules can occur during different stages of the microtubule lifecycle, such as nucleation or elongation. It can take place at either end of the microtubule or in the microtubule lattices as a whole. The lifespan of individual microtubules within a cell varies according to the cell type and stage of the cell cycle. During interphase, the lifespan of the microtubule is about 30 minutes, while during cell division, it is about 15 minutes. In axonal microtubules of...
2.7K