Thesis

Fanconi Anemia Complementary Group I Gene is Regulated by Sp1 and Elk-1 Transcription Factor Binding SItes

Fanconi Anemia Complementary Group I Gene Is Regulated by SP1 and Elk-1 Transcription Factor Binding Sites By Gavin Long Fanconi Anemia Complementary Group I gene (FANCI) plays a role in disorders affecting bone marrow, resulting in anemia, leukopenia, thrombopenia, chromosome instability, and susceptibility to several types of cancer. The FANCI gene produces a protein crucial to the structure and function of a protein complex that repairs double-strand breaks in DNA and interstrand DNA cross-links. The putative promoter region of the human FANCI gene was identified and amplified from genomic DNA. Progressive truncations of the putative promoter were paired with a luciferase reporter vector and assayed for relative gene expression. The greatest relative expression indicates a positive regulatory element(s) likely to drive transcription between the -70bp and +52bp region flanking the Genbank-assigned transcription start site (TSS). Three TSS were experimentally found by 5'RACE thirty or more base pairs downstream of the Genbank-assigned TSS. Computer modeling predicted multiple consensus transcription factor binding sites including Sp1 and ELK-1 within this region, and these were found to be conserved through evolution via multiple sequence alignment of nineteen species. The consensus transcription factor binding sites predicted and evolutionarily conserved were specifically deleted using PCR by exclusion, and relative luciferase activity identified the sequences contributing to the regulation of the FANCI gene. Taken together, these results indicate the FANCI gene is regulated by a dispersed promoter containing two Sp1 sites and an Elk-1 site essential for maximal transcriptional expression. Understanding dysregulation patterns of FANCI can help to develop better treatments and cures for diseases related to Dysregulation of the FANCI gene.

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