title: Two duplicated genes DDI2 and DDI3 in budding yeast encode a cyanamide hydratase and are induced by cyanamide.
abstract: Two DNA damage-inducible genes in Saccharomyces cerevisiae, DDI2 and DDI3, are identical and encode putative HD domain-containing proteins, whose functions are currently unknown. Because Ddi2/3 also shows limited homology to a fungal cyanamide hydratase that converts cyanamide to urea, we tested the enzymatic activity of recombinant Ddi2. To this end, we developed a novel enzymatic assay and determined that the Km value of the recombinant Ddi2/3 for cyanamide is 17.3 ± 0.05 mm, and its activity requires conserved residues in the HD domain. Unlike most other DNA damage-inducible genes, DDI2/3 is only induced by a specific set of alkylating agents and surprisingly is strongly induced by cyanamide. To characterize the biological function of DDI2/3, we sequentially deleted both DDI genes and found that the double mutant was unable to metabolize cyanamide and became much more sensitive to growth inhibition by cyanamide, suggesting that the DDI2/3 genes protect host cells from cyanamide toxicity. Despite the physiological relevance of the cyanamide induction, DDI2/3 is not involved in its own transcriptional regulation. The significance of cyanamide hydratase activity and its induced expression is discussed.
authors.affiliation: From the Departments of Microbiology and Immunology and.
authors.forename: Jia
authors.initials: J
authors.lastname: Li
authors.affiliation: From the Departments of Microbiology and Immunology and.
authors.forename: Michael
authors.initials: M
authors.lastname: Biss
authors.affiliation: From the Departments of Microbiology and Immunology and.
authors.forename: Yu
authors.initials: Y
authors.lastname: Fu
authors.affiliation: the College of Life Sciences, Capital Normal University, Beijing 100048, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.affiliation: Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada and.
authors.forename: Stanley A
authors.initials: SA
authors.lastname: Moore
authors.affiliation: From the Departments of Microbiology and Immunology and the College of Life Sciences, Capital Normal University, Beijing 100048, China wei.xiao@usask.ca.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiao
mesh_terms: D003484:Cyanamide / Q000378:metabolism / Q000494:pharmacology; D004790:Enzyme Induction / Q000187:drug effects; D017353:Gene Deletion; D020440:Gene Duplication / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects / Q000502:physiology*; D006836:Hydro-Lyases / Q000096:biosynthesis* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics; D014508:Urea / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D003484:Cyanamide; D014508:Urea; D006836:Hydro-Lyases
keywords: HD domain; cyanamide hydratase; enzyme; gene duplication; gene regulation; hydratase; metabolism; transcription; transcriptional regulation; yeast
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