Filaments and fingers: Novel structural aspects of the single septin from Chlamydomonas reinhardtii

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Pinto, Andressa P. A. ; Pereira, Humberto M. ; Zeraik, Ana E. ; Ciol, Heloisa ; Ferreira, Frederico M. ; Brandao-Neto, Jose ; DeMarco, Ricardo ; Navarro, Marcos V. A. S. ; Risi, Cristina ; Galkin, Vitold E. ; Garratt, Richard C. ; Araujo, Ana P. U.

• Tipo de documento: Artigo Científico
• Fonte: Journal of Biological Chemistry; v. 292, n. 26, p. 10899-10911, JUN 30 2017.
• Citações Web of Science: 2

Resumo

Septins are filament-forming GTP-binding proteins involved in many essential cellular events related to cytoskeletal dynamics and maintenance. Septins can self-assemble into heterocomplexes, which polymerize into highly organized, cell membrane-interacting filaments. The number of septin genes varies among organisms, and although their structure and function have been thoroughly studied in opisthokonts (including animals and fungi), no structural studies have been reported for other organisms. This makes the single septin from Chlamydomonas (CrSEPT) a particularly attractive model for investigating whether functional homopolymeric septin filaments also exist. CrSEPT was detected at the base of the flagella in Chlamydomonas, suggesting that CrSEPT is involved in the formation of a membrane-diffusion barrier. Using transmission electron microscopy, we observed that recombinant CrSEPT forms long filaments with dimensions comparable with those of the canonical structure described for opisthokonts. The GTP-binding domain of CrSEPT purified as a nucleotide-free monomer that hydrolyzes GTP and readily binds its analog guanosine 5'-3-O-(thio) triphosphate. We also found that upon nucleotide binding, CrSEPT formed dimers that were stabilized by an interface involving the ligand (G-interface). Across this interface, one monomer supplied a catalytic arginine to the opposing subunit, greatly accelerating the rate of GTP hydrolysis. This is the first report of an arginine finger observed in a septin and suggests that CrSEPT may act as its own GTP-activating protein. The finger is conserved in all algal septin sequences, suggesting a possible correlation between the ability to form homopolymeric filaments and the accelerated rate of hydrolysis that it provides. (AU)

• Processo FAPESP: 11/10152-7 - Caracterização estrutural e funcional da septina de Chlamydomonas reinhardtii
• Beneficiário: Ana Paula Ulian de Araujo
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 14/15546-1 - Septinas: estudos comparativos visando correlacionar estrutura e função
• Beneficiário: Richard Charles Garratt
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 12/21259-0 - Septina de Chlamydomonas reinhardtii: estudos com foco em sua expressão e função
• Beneficiário: Heloisa Ciol
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 12/14223-9 - Estudos estruturais e cinéticos das enzimas da via de salvação de purina de Schistosoma mansoni
• Beneficiário: Humberto D'Muniz Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 13/20715-4 - Estudo da correlação entre a dinâmica de septinas e a homeostase de Ca2+ em células musculares de Schistosoma mansoni
• Beneficiário: Ana Eliza Zeraik
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado