Asparagine residue in the rho Gene Product Is the Modification Site for Botulinum ADP-ribosyltransferase

Sekine, Akihiro; Fujiwara, Motohatsu; Narumiya, Shuh

doi:10.1016/s0021-9258(18)81834-8

Cited by 523 publications

(64 citation statements)

References 25 publications

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“…Microinjection of C3 transferase could be labeled with rhodamine-conjugated streptavidin. Incompletely or noningested Baf-3 cells were therefore from Clostridium botulinum which ADP ribosylates and inactivates Rho [15] significantly increased apoptotic cell identified by their red fluorescence (see arrowheads). uptake.…”

Section: Figurementioning

confidence: 99%

Requirement for Rho GTPases and PI 3-kinases during apoptotic cell phagocytosis by macrophages

2001

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In vivo, apoptotic cells are removed by surrounding phagocytes, a process thought to be essential for tissue remodeling and the resolution of inflammation [1]. Although apoptotic cells are known to be efficiently phagocytosed by macrophages, the mechanisms whereby their interaction with the phagocytes triggers their engulfment have not been described in mammals. Here, we report that primary murine bone marrow-derived macrophages (using alpha(v)beta(3) integrin for apoptotic cell uptake) extend lamellipodia to engulf apoptotic cells and form an actin cup where phosphotyrosine accumulates. Rho GTPases and PI 3-kinases have been widely implicated in the regulation of the actin cytoskeleton [2, 3]. We show that inhibition of Rho GTPases by Clostridium difficile toxin B prevents apoptotic cell phagocytosis and inhibits the accumulation of both F-actin and phosphotyrosine. Importantly, the Rho GTPases Rac1 and Cdc42 are required for apoptotic cell uptake whereas Rho inhibition enhances uptake. The PI 3-kinase inhibitor LY294002 also prevents apoptotic cell phagocytosis but has no effect on the accumulation of F actin and phosphotyrosine. These results indicate that both Rho GTPases and PI 3-kinases are involved in apoptotic cell phagocytosis but that they play distinct roles in this process.

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Section: Figurementioning

confidence: 99%

Requirement for Rho GTPases and PI 3-kinases during apoptotic cell phagocytosis by macrophages

2001

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“…According to amino acid sequence similarity, yeast and animal Rho-type GTPases are categorized into at least three subfamilies, CDC42, Rac, and Rho (Chardin, 1993). The C3 exoenzyme from Clostridium botulinum has been shown to preferentially ADP-ribosylate and thus to inactivate the Rho subfamily (Aktories et al, 1989;Sekine et al, 1989;Sugai et al, 1992). Thus, the C3 toxin is useful for distinguishing the biological function for the Rho subfamily from other subfamilies (Chardin et al, 1989;Sugai et al, 1992;Takaishi et al, 1993).…”

Section: Distinct Growth Behavior 1s Lnduced By the C3 Toxin A Rho-specific Adp-ribosyltransferasementioning

confidence: 99%

Inhibition of Pollen Tube Elongation by Microinjected Anti-Rop1Ps Antibodies Suggests a Crucial Role for Rho-Type GTPases in the Control of Tip Growth

Lin¹,

Yang²

1997

The Plant Cell

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Microinjection of anti-Rop1Ps antibodies was used to assess the function of a tip-localized Rho-type GTPase, Rop, in controlling pollen tube growth. Injected antibodies induced sustained growth arrest within 1 to 2 min after injection but did not affect cytoplasmic streaming. Coinjection with Rop rescued antibody-induced growth inhibition, indicating that injected antibodies specifically block the activity of Rop GTPases. Antibody-induced inhibition was significantly enhanced in the presence of a lower threshold of extracellular [Ca2+] or a subinhibitory dosage of caffeine. In contrast, injection of the C3 toxin, which inactivates a different Rho-type GTPase, arrested tube elongation 10 to 20 min after injection. C3-induced growth arrest was accompanied by the cessation of cytoplasmic streaming. These data suggest that Rho-type GTPases play a pivotal role in the control of pollen tube elongation. We propose that Rop may regulate a Ca2+-dependent pathway involved in vesicle docking/fusion, whereas a C3-sensitive Rho GTPase may mediate cytoplasmic streaming.

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“…ADPr modifies proteins at very low levels, making enrichment methods necessary. MS-based analysis is further complicated by the dynamic nature of ADPr, with a rapid enzymatic turnover of the modification (Alvarez- Gonzalez and Althaus, 1989), numerous modifiable amino acid residues (Altmeyer et al, 2009;Bonfiglio et al, 2017b;McDonald and Moss, 1994;Moss and Vaughan, 1977;Ogata et al, 1980;Sekine et al, 1989;Van Ness et al, 1980), and the highly labile nature of the modification (Bonfiglio et al, 2017a).…”

Section: Introductionmentioning

confidence: 99%

Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation

et al. 2020

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ADP-ribosylation (ADPr) is a post-translational modification that plays pivotal roles in a wide range of cellular processes. Mass spectrometry (MS)-based analysis of ADPr under physiological conditions, without relying on genetic or chemical perturbation, has been hindered by technical limitations. Here, we describe the applicability of activated ion electron transfer dissociation (AI-ETD) for MS-based proteomics analysis of physiological ADPr using our unbiased Af1521 enrichment strategy. To benchmark AI-ETD, we profile 9,000 ADPr peptides mapping to >5,000 unique ADPr sites from a limited number of cells exposed to oxidative stress and identify 120% and 28% more ADPr peptides compared to contemporary strategies using ETD and electron-transfer higher-energy collisional dissociation (EThcD), respectively. Under physiological conditions, AI-ETD identifies 450 ADPr sites on low-abundant proteins, including in vivo cysteine modifications on poly(ADP-ribosyl)polymerase (PARP) 8 and tyrosine modifications on PARP14, hinting at specialist enzymatic functions for these enzymes. Collectively, our data provide insights into the physiological regulation of ADPr.

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Asparagine residue in the rho Gene Product Is the Modification Site for Botulinum ADP-ribosyltransferase

Cited by 523 publications

References 25 publications

Requirement for Rho GTPases and PI 3-kinases during apoptotic cell phagocytosis by macrophages

Requirement for Rho GTPases and PI 3-kinases during apoptotic cell phagocytosis by macrophages

Inhibition of Pollen Tube Elongation by Microinjected Anti-Rop1Ps Antibodies Suggests a Crucial Role for Rho-Type GTPases in the Control of Tip Growth

Mapping Physiological ADP-Ribosylation Using Activated Ion Electron Transfer Dissociation

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