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Transcriptional regulation of the human neutral ceramidase gene.

Abstract：

:Ceramidases play a critical role in generating sphingosine-1-phosphate by hydrolyzing ceramide into sphingosine, a substrate for sphingosine kinase. In order to elucidate its transcriptional regulation, we identify here a putative promoter region in the 5'-UTR of the human neutral CDase (nCDase) gene. Using human genomic DNA, we cloned a 3000 bp region upstream of the translational start site of the nCDase gene. Luciferase reporter analyses demonstrated that this 3000 bp region had promoter activity, with the strongest induction occurring within the first 200 bp. Computational analysis revealed the 200 bp essential promoter region contained several well-characterized promoter elements, lacked a conical TATA box, but did contain a reverse oriented CCAAT box, a feature common to housekeeping genes. Electrophoretic mobility shift assays demonstrated that the identified candidate transcriptional response elements (TRE) bind their respective transcription factors, including NF-Y, AP-2, Oct-1, and GATA. Mutagenic analyses of the TRE revealed that these sites regulated promoter activity and mutating an individual site decreased promoter reporter activity by up to 50%. Together, our findings suggest that regulation of nCDase expression involves coordinated TATA-less transcriptional activity.

journal_name

Arch Biochem Biophys

journal_title

Archives of biochemistry and biophysics

authors

O'Neill SM,Yun JK,Fox TE,Kester M

doi

10.1016/j.abb.2011.04.012

subject

Has Abstract

pub_date

2011-07-01 00:00:00

pages

21-30

issue

1-2

eissn

0003-9861

issn

1096-0384

pii

S0003-9861(11)00153-6

journal_volume

511

pub_type

杂志文章

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