International DNA Day and Genome Congress, Kırşehir, Turkey, 24 - 28 April 2017, vol.1, no.1, pp.1
Aims and Scopes: Trehalose, acummulated under stress conditions in Saccharomyces cerevisiae yeast strains, is hydrolised by neutral trehalase (Nth 1p), when enviromental conditions return to normal circumstances . Nth1p is encoded by NTH1 gene which is 2256bp in length and is not include intron [1,2]. Tec1p, which is a transcription factor, is encoded by TEC1 gene (Transcription Enhancement Control 1) and it has increased chronological lifespan owing to the MAPK (Mitogen-Activated Protein Kinase) and TORC1 (Target of Rapamycin) pathways [3,4]. Tec1p has controlled transcripts by binding to genes, contained TAE/ATTS elements ((A(G/C/T)C(G)AT(A)TC(G)C(G)C(T)C(T/A/G)) in promotor region. Transcriptional regulation of NTH1 gene is carried out by TOR signal system and NTH1 gene include TEA/ATTS elements, in order to bind Tec1p on NTH1 promotor region. In our study, therefore, in silico binding model of TEC1 Transcription Factor on NTH1 Promotor was done.
Materials and Methods: In this research, NTH1 promotor region (1000 bp in lenght) downloaded from SGD ( Saccharomyces cerevisiae Genome Database) was used. Tec1p TAE/ATTS DNA binding site was detected on the NTH1 promotor by using JASPAR CORE database. Predicted-3D modelling of whole aminoacid sequences (486 aminoacids) of Tec1 protein and aminoacid sequences which include TEA/ATTS DNA binding site (75 aminoacids) was carried out using I-TASSER software. Tec 1p binding elements in NTH1 promotor were turned into pdb format sending to 3D-DART webserver tool. Pdb files, obtained from I-TASSER software and 3D-DART webserver, were sent to NPDock-Genesilico.pl websites and predicted binding model results were attained. The obtained models were visualized by using VMD (Visual Moleculaer Dynamics) program.
Results and Discussion: It was determined that there is high possibility of binding of Tec1 transcription factor to NTH1 promotor, owing to in silico DNA-TF binding modelling which is carried out by using databases and computer tools. In vivo studies which supported 3D model related to NTH1 transcriptional regulation on the Δtec1 mutants have been proceeding.