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.

• Document type: Journal article
• Source: Journal of Biological Chemistry; v. 292, n. 26, p. 10899-10911, JUN 30 2017.
• Web of Science Citations: 2

Abstract

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)

• FAPESP's process: 11/10152-7 - Structural and functional characterization of Chlamydomonas reinhardtii septin
• Grantee: Ana Paula Ulian de Araujo
• Support Opportunities: Regular Research Grants
• FAPESP's process: 14/15546-1 - Septins: comparative studies and the correlation between structure and function
• Grantee: Richard Charles Garratt
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/21259-0 - Septin of Chlamydomonas reinhardtii: studies focusing its expression and function
• Grantee: Heloisa Ciol
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 12/14223-9 - Structural and kinetical studies of the enzymes involved in the purine metabolism in Schistosoma mansoni
• Grantee: Humberto D'Muniz Pereira
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/20715-4 - Studies of the correlation between septin dynamics and Ca2+ homeostasis in muscular cells from Schistosoma mansoni
• Grantee: Ana Eliza Zeraik
• Support Opportunities: Scholarships in Brazil - Post-Doctoral