C. Ciências Biológicas - 3. Bioquímica - 1. Biologia Molecular
HUMAN SEPTINS 6 AND 8 INTERACT WITH SEVERAL OTHER SEPTINS AND WITH PROTEINS INVOLVED IN SIGNALING AND TRANSCRIPTIONAL REGULATION
Marcel Nakahira1, 2
Maria Eugênia Ribeiro de Camargo1
Marcos Rodrigo Alborghetti1, 2
Eliana Maria Assmann1
Jörg Kobarg1, 2, 3
1. Laboratório Nacional de Luz Síncrotron - LNLS
2. Universidade Estadual de Campinas - UNICAMP
3. PhD - Laboratório de Biologia Molecular - Supervisor
INTRODUÇÃO:Septins are a conserved family of GTP binding protein that were first identified in yeast. However, they are also now known to be widely expressed in other fungi and metazoa. The newest mammalian member of this growing family of proteins is septin 14, which has been recently isolated and characterized from human testis. Typically, septins such as those studied here, are 40- to 60-kDa GTPases that assemble to filamentous fibres that may serve as scaffolds in diverse cellular functions, including membrane dynamics, neuronal polarity, vesicle trafficking, apoptosis, and cytoskeletal remodeling. Furthermore, there is growing evidence that they may also play important roles in some human pathologies, including Alzheimer’s disease, Parkinson’s disease and cancer. However, their detailed structural properties and modes of assembly and action have been only recently begun to be addressed. Here, we report our preliminary results on the attempt to identify proteins that interact with the human septins 6 and 8.METODOLOGIA:PCR primers were designed and synthesized according to the SEPTIN 6 and 8 cDNA sequence. The PCR amplification was performed with a Human cDNA library as a template. The PCR product was sub-cloned into pBTM116 vector. The yeast two-hybrid system (Y2HS) was used to screen both human fetal brain and human leukocyte cDNA libraries, using septins 6 and 8 pBTM116 vector, as “baits”.RESULTADOS:Septin 6 has been shown to interact with more different proteins than any other septin so far tested in the Y2HS. A recent report had shown that septin 6 forms a complex with septin 2 and 7, where it is “sandwiched” between the latter two. This may indicate that it has a greater propensity to interact with other proteins. The results of two-hybrid system screen show interesting proteins such as several other septins (including septins 1, 2, 4, 5, 7 and principally 9) and other proteins related to signaling processes and the regulation of transcription.CONCLUSÕES:These data show that septins 6 and 8 interact with other septins as well as variety other proteins, indicating many play important roles in the cell functions.
Instituição de fomento: CAPES, FAPESP, CNPq and LNLS
Palavras-chave: GTPase, Septin, Yeast Two Hybrid System
E-mail para contato: jkobarg@lnls.br
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