Molecular cloning, modeling and differential expression of a gene encoding a silent information regulator-like protein from Sporothrix schenckii

Abstract

Sporothrix schenckii (S. schenckii) is a dimorphic fungus that produces lymphocutaneous lesions. The signature characteristic of S. schenckii is a temperature-induced phase transition. Silent information regulator (Sir) has been proven to be involved in phenotypic switching in Saccharomyces cerevisiae (S. cerevisiae) and Candida albicans (C. albicans) by organizing chromatin structure. In this study, we isolated and characterized a Sir homologue gene, designated as SsSir2, from the yeast form of S. schenckii. The full-length SsSir2 cDNA sequence is 1753 bp in size and contains an open reading frame of 1329 bp encoding 442 amino acids. The predicted molecular mass of SsSir2 is 48.1 kDa with an estimated theoretical isoelectric point of 4.6. The SsSir2 kinase domain shows a 78% identity with that of Hst2, a Sir2 Ib gene from S. cerevisiae. Three exons and two introns were identified within the 1472-bp SsSir2 genomic DNA sequence of S. schenckii. A three-dimensional model of SsSir2 was constructed using a homology modeling method, and its reliability was evaluated. The active site of SsSir2 was identified by docking simulation, which indicated that several important residues, such as Asn127 and Asp129, play an important role in the histone deacetylase activity of Sir2 family proteins. The differential expression of the SsSir2 in two stages was demonstrated by real-time RT-PCR. The expression of SsSir2 was higher in the yeast stage compared with that in the mycelial one, which indicated that SsSir2 may be involved in the phenotypic switching and morphogenesis of the yeast phase in S. schenckii.

The Sporothrix schenckii Gene Encoding for the Ribosomal Protein L6 Has Constitutive and Stable Expression and Works as an Endogenous Control in Gene Expression Analysis

“…Gene silencing, disruption, edition or over expression are usually accompanied by analysis of gene expression, which rely on data normalization using endogenous genes with stable and constitutive expression across different growth conditions. Previous works in S. schenckii have used the gene encoding for β-tubulin ( Hu et al, 2015 ), ribosomal protein L34 ( Teixeira et al, 2015 ), and 18S ribosomal RNA ( de Jesús-Berríos and Rodríguez-del Valle, 2002 ; Hou et al, 2013 , 2014 ; Zhang et al, 2013 ). However, the expression of these genes is assumed to be stable and constitutive.…”

Will Sirtuin 2 Be a Promising Target for Neuroinflammatory Disorders?

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Saccharomyces cerevisiae as a skin physiology, pathology, and treatment model

“…This marker shares significant homology with the Sir2 gene, which is involved in phenotypic switching in S. cerevisiae. In this way, initial findings from yeast were used to find a related marker in a different organism [4]. Another group used S. cerevisiae to study the role of GPR18, a GPCR protein that was abundantly overexpressed in melanoma metastases.…”