Rac1 regulates heat shock responses by reorganization of vimentin filaments: Identification using MALDI-TOF MS

Lee, S. Y.; Song, Eun Joo; Kim, H. J.; Kang, Hyojung; Kim, J. H.; Lee, K. J.

doi:10.1038/sj.cdd.4400923

Cited by 15 publications

(12 citation statements)

References 32 publications

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“…The results presented here show extensive reorganization of the intermediate filament network, consisting in a perinuclear collapse of vimentin filaments. It is interesting that a similar collapse of the vimentin network has been reported in several cell types that underwent a heat-shock response (39,40); however, this is the first report of this kind of reorganization in MC. These changes could constitute part of the defense mechanisms triggered by 15d-PGJ 2 in MC.…”

Section: Discussionmentioning

confidence: 84%

Identification of Novel Protein Targets for Modification by 15-Deoxy-Δ12,14-Prostaglandin J2 in Mesangial Cells Reveals Multiple Interactions with the Cytoskeleton

Stamatakis¹,

Sánchez‐Gómez²

2006

Journal of the American Society of Nephrology

108

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The cyclopentenone prostaglandin 15-deoxy-⌬ 12,14 -PGJ 2 (15d-PGJ 2 ) has been shown to display protective effects against renal injury or inflammation. In cultured mesangial cells (MC), 15d-PGJ 2 inhibits the expression of proinflammatory genes and modulates cell proliferation. Therefore, cyclopentenone prostaglandins (cyPG) have been envisaged as a promise in the treatment of renal disease. The effects of 15d-PGJ 2 may be dependent on or independent from its role as a peroxisome proliferator-activated receptor agonist. It was shown recently that an important determinant for the peroxisome proliferatoractivated receptor-independent effects of 15d-PGJ 2 is the capacity to modify proteins covalently and alter their function. However, a limited number of protein targets have been identified to date. Herein is shown that a biotinylated derivative of 15d-PGJ 2 recapitulates the effects of 15d-PGJ 2 on the stress response and inhibition of inducible nitric oxide synthase levels and forms stable adducts with proteins in intact MC. Biotinylated 15d-PGJ 2 was then used to identify proteins that potentially are involved in cyPG biologic effects. Extracts from biotinylated 15d-PGJ 2 -treated MC were separated by two-dimensional electrophoresis, and the spots of interest were analyzed by mass spectrometry. Identified targets include proteins that are regulated by oxidative stress, such as heat-shock protein 90 and nucleoside diphosphate kinase, as well as proteins that are involved in cytoskeletal organization, such as actin, tubulin, vimentin, and tropomyosin. Biotinylated 15d-PGJ 2 binding to several targets was confirmed by avidin pull-down. Consistent with these findings, 15d-PGJ 2 induced early reorganization of vimentin and tubulin in MC. The cyclopentenone moiety and the presence of cysteine were important for vimentin rearrangement. These studies may contribute to the understanding of the mechanism of action and therapeutic potential of cyPG.

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Section: Discussionmentioning

confidence: 84%

Identification of Novel Protein Targets for Modification by 15-Deoxy-Δ12,14-Prostaglandin J2 in Mesangial Cells Reveals Multiple Interactions with the Cytoskeleton

Stamatakis¹,

Sánchez‐Gómez²

2006

Journal of the American Society of Nephrology

108

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“…In support of this, it has been previously demonstrated that alterations of cytoskeleton components dynamics affect cell migration [75]. Notably, Rac1 and vimentin functions are connected [51][52][53][54]. Indeed, Rac1 regulates reorganization of vimentin filaments, as Rac1 activation causes phosphorylation of vimentin at Ser38 inducing vimentin filament disassembly [52][53][54].…”

Section: Discussionmentioning

confidence: 87%

“…In fact, vimentin regulates Rac1 activation while activation of Rac1 induces vimentin phosphorylation and disassembly [51][52][53][54]. Furthermore, we have previously shown that Rab7a interacts with vimentin and that depletion of Rab7a determines a lower phosphorylated state of the head domain of vimentin and a higher abundance of vimentin in the insoluble, filamentous fraction [17].…”

Section: Rab7a Regulates Vimentin Filaments Orientation During Cell Mmentioning

confidence: 99%

Rab7a regulates cell migration through Rac1 and vimentin

Margiotta

Progida

Bakke

et al. 2017

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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Rab7a, a small GTPase of the Rab family, is localized to late endosomes and controls late endocytic trafficking. The discovery of several Rab7a interacting proteins revealed that Rab7a function is closely connected to cytoskeletal elements. Indeed, Rab7a recruits on vesicles RILP and FYCO that are responsible for the movement of Rab7a-positive vesicles and/or organelles on microtubule tracks, but also directly interacts with Rac1, a fundamental regulator of actin cytoskeleton, and with peripherin and vimentin, two intermediate filament proteins. Considering all these interactions and, in particular, the fact that Rac1 and vimentin are key factors for cellular motility, we investigated a possible role of Rab7a in cell migration. We show here that Rab7a is needed for cell migration as Rab7a depletion causes slower migration of NCI H1299 cells affecting cell velocity and directness.Rab7a depletion negatively affects adhesion and spreading onto fibronectin substrates, altering β1-integrin activation, localization and intracellular trafficking, and myosin X localization. In fact, Rab7a-depleted cells show 40% less filopodia and active integrin accumulates at the leading edge of migrating cells. Furthermore, Rab7a depletion decreases the amount of active Rac1 but not its abundance and reduces the number of cells with vimentin filaments facing the wound, indicating that Rab7a has a role in the orientation of vimentin filaments during migration. In conclusion, our results demonstrate a key role of Rab7a in the regulation of different aspects of cell migration.

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“…This is consistent with the finding that heat shock induces protein synthesis blockage and recovery (28), and cytoskeletal protein collapse and recovery (48) during recovery after heat shock. Intermediate filament vimentin modifications were identified in response to heat shock previously (48). Translocations of vimentin by various stresses were reported (49,50).…”

Section: Proteins That Act In Energy Metabolismmentioning

confidence: 99%

Proteomic Analysis of Protein Phosphorylations in Heat Shock Response and Thermotolerance

Kim¹,

Song²,

Lee

2002

Journal of Biological Chemistry

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Heat shock (HS) induces a wide variety of biological processes, including inhibition of protein synthesis, elevated expression of heat shock proteins, induction of thermotolerance, and apoptotic cell death in a dose-dependent manner. We compared phosphorylated proteins in heat-shocked and thermotolerant cells using proteome analysis. After HS treatment of control RIF-1 and their thermotolerant derivatives, TR-RIF-1 cells, cellular proteins were separated by two-dimensional gel electrophoresis and the phosphorylated proteins were detected with the anti-phosphotyrosine antibodies. We found that 93 proteins showed significant changes in phosphorylation between control and thermotolerant cells as a function of recovery time after HS; we identified 81 of these proteins with peptide mass fingerprinting using MALDI-TOF MS after in-gel trypsin digestion. These phosphorylated proteins exhibit various cellular functions, including chaperones, ion channels, signaling molecules, in transcription and translation processes, in amino acid biosynthesis, oxidoreduction, energy metabolism, and cell motility or structure, suggesting that HS turns on the various signaling pathways by activating protein-tyrosine kinases (PTKs). Of these, 20 proteins were previously identified phosphorylated proteins and 64 were newly identified. These proteins can be grouped into three families: 1) proteins highly phosphorylated in TR-RIF-1 cells at basal level and phosphorylated more significantly by HS in RIF-1 than TR-RIF-1; 2) proteins highly phosphorylated in control RIF-1 cells at basal level and phosphorylated more easily by HS in TR-RIF-1 than in RIF-1 cells; and 3) proteins with a similar basal phosphorylation level in both RIF-1 and TR-RIF-1 cells and responding to HS similarly in both cells. Most of the phosphorylated proteins are presumably involved in HS signaling in different ways, with the first and second families of proteins influencing thermotolerance. The possible tyrosine phosphorylation sites, the possible PTKs phosphorylating these proteins, and the proteins binding to these phosphorylated sites were predicted by the Netphos, ScanProsite, and Scansite programs. These results suggest that HS can activate various PTKs and HS responses can be regulated by phosphorylations of proteins having various functions.Heat shock responses are well conserved phenomena through evolution. Modest elevations of temperature induce apoptotic cell death. A common feature of the heat shock response is that an initial, nonlethal heat shock provides a transient resistance against subsequent lethal heat shock. This phenomenon is called thermotolerance. Thermotolerant cells induce the overexpression of a family of heat shock proteins (Hsps) 1 and are thereby protected from cell death caused by various stresses. This suggests that the chaperonic function of Hsps is associated with the development of thermotolerance. However, the details of the molecular events underlying heat shock responses are not well defined.Heat shock causes a dramatic reprogramming...

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Rac1 regulates heat shock responses by reorganization of vimentin filaments: Identification using MALDI-TOF MS

Cited by 15 publications

References 32 publications

Identification of Novel Protein Targets for Modification by 15-Deoxy-Δ12,14-Prostaglandin J2 in Mesangial Cells Reveals Multiple Interactions with the Cytoskeleton

Identification of Novel Protein Targets for Modification by 15-Deoxy-Δ12,14-Prostaglandin J2 in Mesangial Cells Reveals Multiple Interactions with the Cytoskeleton

Rab7a regulates cell migration through Rac1 and vimentin

Proteomic Analysis of Protein Phosphorylations in Heat Shock Response and Thermotolerance

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