Mdecular cloning and analysis of S' -flanking region of human MYPT1 gene

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MYPT1, a subunit of myosin phosphatase, has critical functions including activation and regulation of phosphatase activity.We have cloned and sequenced a 5 kb genomic fragment in the 5'-flanking region of the human MYPT1 gene. The transcription initiation site (+1) was 268 bp upstream from the translation start site. In this promoter there are no canonical TATA or CAAT box elements but there is a high GC-rich sequence (-58 to -64). Basal promoter activity was due to the GC-rich region that contained one Sp 1 transcription factor-binding site. The specific interaction of Sp 1 with this region was demonstrated by the electrophoresis mobility shift assay. Therefore, it was concluded that the MYPT1 gene is a housekeeping gene. Luciferase reporter assays showed the presence of two regions for positive elements (-246 to -230 and -170 to -198) and one for a negative element (-199 to -229).