Project description:ChIP-seq analysis was used to identify B. dermatitidis genes bound by the GATA transcription factor encoded by SREB during growth as yeast at 37oC SREB was engineered to contain an in-frame 3x-hemagglutinin (HA) epitope tag at the C-terminus. The SREB-3xHA construct was under control of its native promoter and contained the 3-untranslated region. Using Agrobacterium tumefaciens, B. dermatitidis ATCC 26199 was transformed with the SREB-3xHA construct (referred to a SREB-3xHA strain in this document). The SREB-3xHA construct was functional because retransformation of SREB? with the construct complented the null mutant. Chromatin was extracted and sheared from ATCC 26199 and SREB-3xHA yeast grown in liquid Histoplasma macrophage medium (HMM) containing 10 ?M iron sulfate (FeSO4) at 37oC. ATCC 26199 was the untagged control strain.
Project description:ChIP-seq analysis was used to identify B. dermatitidis genes bound by the GATA transcription factor encoded by SREB during growth as yeast at 37oC
Project description:The goal of this study was to capture the transcriptional response regulated by the GATA transcription factor encoded by SREB during the first 48-hours of the temperature-dependent shift from yeast (37oC) to mycelia (22oC). Gene expression microarrays were used to compare an SREB null mutant (SREBΔ) to an isogenic wild type strain (ATCC 26199). SREB null mutants fail to fully complete the conversion from yeast to sporulating mycelia after a drop in temperature from 37oC to 22oC, and under iron-replete conditions cannot properly repress siderophore biosynthesis. Deletion of SREB in ATCC 26199 produced SREBΔ. The isogenic wild-type strain is ATCC 26199.
