Structure of the gene encoding dihydrolipoyl transacylase (E2) component of human branched chain α-keto acid dehydrogenase complex and characterization of an E2 pseudogene

Kim S. Lau, W. Joseph Herring, Jacinta L. Chuang, Martha McKean, Dean J. Danner, Rody P. Cox, David T. Chuang

Research output: Contribution to journalArticle

Abstract

We have determined the structural organization of the dihydrolipoyl transacylase (E2) gene of the human branched chain α-keto acid dehydrogenase complex. The single copy E2 gene spans approximately 68 kilobases of genomic DNA. The complete coding region consisting of the 5′- and 3′-untranslated regions, the mitochondrial targeting sequence (61 amino acids), and the mature E2 sequence (421 amino acids) are encoded by 11 exons ranging from 62 to 2239 base pairs. All the donor and acceptor splice sites conform to the gt-ag rule. Sequence analysis of the promoter-regulatory region showed the presence of a "CAAT box"-like sequence 537 bases upstream of the transcription initiation site. The "TATA box"-like sequence is absent. Also located in this region are sequences resembling glucocorticoid-responsive and cAMF-responsive elements, fat-specific elements, and Sp1- and AP-2-binding sites. Several sets of direct and inverted repeats are also present. Promoter assays using human hepatoma cells (Hep-G2) and Swiss mouse preadipocytes (3T3-L1) showed that a 4.1-kilobase PstI fragment upstream of the transcription start site confers high expression of the luciferase reporter gene. Moreover, an intronless E2 pseudogene was isolated. It corresponds to the complete mitochondrial presequence and the lipoyl-bearing domain that are encoded by exons I through IV of the functional E2 gene. However ,the E2 pseudogene contains multiple base changes, deletions, and insertions, and is flanked by short direct repeats. The data indicate that the E2 pseudogene is a retroposon.

Original languageEnglish (US)
Pages (from-to)24090-24096
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number33
StatePublished - Nov 25 1992
Externally publishedYes

Fingerprint

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Pseudogenes
Gene encoding
RNA Splice Sites
Genes
Nucleic Acid Repetitive Sequences
Transcription Initiation Site
Amino Acid Sequence
Exons
Retroelements
TATA Box
Bearings (structural)
Nucleic Acid Regulatory Sequences
5' Untranslated Regions
Hep G2 Cells
3' Untranslated Regions
Luciferases
Reporter Genes
Amino Acids
Genetic Promoter Regions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structure of the gene encoding dihydrolipoyl transacylase (E2) component of human branched chain α-keto acid dehydrogenase complex and characterization of an E2 pseudogene. / Lau, Kim S.; Herring, W. Joseph; Chuang, Jacinta L.; McKean, Martha; Danner, Dean J.; Cox, Rody P.; Chuang, David T.

In: Journal of Biological Chemistry, Vol. 267, No. 33, 25.11.1992, p. 24090-24096.

Research output: Contribution to journalArticle

Lau, Kim S. ; Herring, W. Joseph ; Chuang, Jacinta L. ; McKean, Martha ; Danner, Dean J. ; Cox, Rody P. ; Chuang, David T. / Structure of the gene encoding dihydrolipoyl transacylase (E2) component of human branched chain α-keto acid dehydrogenase complex and characterization of an E2 pseudogene. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 33. pp. 24090-24096.
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