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The Ras Gene02:38

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The Ras-gene-encoded proteins are regulators of signaling pathways controlling cell proliferation, differentiation, or cell survival. The Ras-gene family in humans constitutes three primary members—the HRas, NRas, and KRas. These genes code for four functionally distinct yet closely related proteins—the HRas, NRas, KRas4A, and KRas4B. The involvement of mutant Ras genes in human cancer was first discovered in 1982 and is among the most common causes of human tumorigenesis.
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Ras and Rho are small monomeric GTPases that act downstream of receptor tyrosine kinase (RTK) and regulate various cellular processes. These GTPases switch between active and inactive states by binding to guanine nucleotides.
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Cell polarity is the asymmetric distribution of cellular and membrane components, making one side of the cell different from the other. This polarity is essential to many processes such as embryogenesis, axon migration, glucose transport across epithelial cells, and directional cell migration. A migrating cell responds to intracellular or extracellular signals via molecular cascades that reorganize the actin cytoskeleton to establish this polarity. In these cells, the Rho family proteins Cdc42,...
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Rab GTPases act in a regulated cascade during membrane fusion, helping the lipid bilayers mix. The Rab family of proteins are active when bound to GTP, and inactive when bound to GDP. Hence, they act as guanine nucleotide-dependent molecular switches. Rab-GTP recognizes and binds to long or short-range tethering proteins to capture the target vesicle. These tethers coordinate with SNAREs on the vesicle and the target membrane to assemble the trans SNARE complex that locks the mixing bilayers.
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Blebs are a type of membrane protrusion formed by the internal hydrostatic pressure of the cytoplasm. Blebs are observed in several cell types, including fibroblasts, immune cells, and single-celled organisms like the amoeba. The primary function of blebs is cell locomotion and apoptosis, but they are also found during necrosis and cell division. The life cycle of a bleb comprises an initiation phase followed by the expansion and retraction phases.
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Excitable Ras dynamics-based screens reveal RasGEFX is required for macropinocytosis and random cell migration.

Koji Iwamoto1, Satomi Matsuoka2,3,4, Masahiro Ueda5,6

  • 1Laboratory of Single Molecule Biology, Graduate School of Science and Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka, 565-0871, Japan.

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Researchers identified key RasGEF proteins in Dictyostelium discoideum that control spontaneous cell migration and macropinocytosis. RasGEFX is crucial for initiating Ras-GTP signaling, essential for cell movement and nutrient uptake.

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Area of Science:

  • Cell Biology
  • Biochemistry
  • Molecular Biology

Background:

  • Eukaryotic cells exhibit chemotaxis, enabling directed movement.
  • Spontaneous signaling of Ras-GTP domains drives random cell migration.
  • The specific molecules generating these spontaneous signals were previously unknown.

Purpose of the Study:

  • To identify the Ras guanine nucleotide exchange factors (RasGEFs) responsible for spontaneous Ras-GTP domain generation in Dictyostelium discoideum.
  • To elucidate the distinct roles of various RasGEFs in regulating cell migration and macropinocytosis.

Main Methods:

  • Live-cell imaging of Ras-GTP spatiotemporal dynamics.
  • Hierarchical clustering analysis of signaling dynamics.
  • Genetic manipulation of RasGEF expression in Dictyostelium discoideum.

Main Results:

  • RasGEFX is essential for the spontaneous generation of Ras-GTP domains and random cell migration in starved cells, working with RasGEFB/M/U.
  • RasGEFX and RasGEFB regulate temporal and spatial aspects of Ras-GTP waves, influencing cell protrusions.
  • RasGEFU and RasGEFM control cell adhesion and migration speed, respectively.
  • RasGEFX uniquely supports Ras/PIP3-driven macropinocytosis in proliferating cells.

Conclusions:

  • Multiple RasGEFs coordinate to control cytoskeletal dynamics for spontaneous motility.
  • RasGEFs play distinct roles in regulating cell migration, adhesion, and macropinocytosis.
  • RasGEFX is a key regulator of both spontaneous migration and macropinocytosis.