Three promoters regulate the transcriptional activity of the human holocarboxylase synthetase gene

Mengna Xia, Sridhar A. Malkaram, Janos Zempleni

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Holocarboxylase synthetase (HLCS) is the only protein biotin ligase in the human proteome. HLCS-dependent biotinylation of carboxylases plays crucial roles in macronutrient metabolism. HLCS appears to be an essential part of multiprotein complexes in the chromatin that cause gene repression and contribute toward genome stability. Consistent with these essential functions, HLCS knockdown causes strong phenotypes including shortened life span and low stress resistance in Drosophila melanogaster, and de-repression of long-terminal repeats in humans, other mammalian cell lines and Drosophila. Despite previous observations that the expression of HLCS depends on biotin status in rats and in human cell lines, little is known about the regulation of HLCS expression. The goal of this study was to identify promoters that regulate the expression of the human HLCS gene. Initially, the human HLCS locus was interrogated in silico using predictors of promoters including sequences of HLCS mRNA and expressed sequence tags, CpG islands, histone marks denoting transcriptionally poised chromatin, transcription factor binding sites and DNaseI hypersensitive regions. Our predictions revealed three putative HLCS promoters, denoted P1, P2 and P3. Promoters lacked a TATA box, which is typical for housekeeping genes. When the three promoters were cloned into a luciferase reporter plasmid, reporter gene activity was at least three times background noise in human breast, colon and kidney cell lines; activities consistently followed the pattern P1>>P3>P2. Promoter activity depended on the concentration of biotin in culture media, but the effect was moderate. We conclude that we have identified promoters in the human HLCS gene.

Original languageEnglish (US)
Pages (from-to)1963-1969
Number of pages7
JournalJournal of Nutritional Biochemistry
Volume24
Issue number11
DOIs
StatePublished - Nov 1 2013

Fingerprint

Human Activities
Genes
Biotin
Cells
Cell Line
Chromatin
holocarboxylase synthetases
Histone Code
Biotinylation
Multiprotein Complexes
TATA Box
CpG Islands
Terminal Repeat Sequences
Genomic Instability
Essential Genes
Expressed Sequence Tags
Proteome
Ligases
Drosophila melanogaster
Luciferases

Keywords

  • Biotin
  • Holocarboxylase synthetase
  • Human
  • Promoter

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Molecular Biology
  • Nutrition and Dietetics
  • Clinical Biochemistry

Cite this

Three promoters regulate the transcriptional activity of the human holocarboxylase synthetase gene. / Xia, Mengna; Malkaram, Sridhar A.; Zempleni, Janos.

In: Journal of Nutritional Biochemistry, Vol. 24, No. 11, 01.11.2013, p. 1963-1969.

Research output: Contribution to journalArticle

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