ZEC, a zinc finger protein with novel binding specificity and transcription regulatory activity

Guoyun Chen, Hisako Muramatsu, Keiko Ichihara-Tanaka, Takashi Muramatsu

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

A novel 114-kDa zinc finger protein, ZEC, has been found by cDNA cloning and characterized. ZEC was strongly expressed in the testis, liver and kidney, and also in embryonic stem cells. Epitope-tagged experiments indicated nuclear localization of ZEC. ZEC contained 18 C2H2 zinc fingers which were organized in two clusters. A ZEC binding DNA sequence, C/GA/TA/TGGTTGGTTGC, which we have designated the GT box, was identified by random oligonucleotide binding selection assay. The GT box did not contain binding sites for other previously characterized transcription factors and thus represented a potentially novel DNA target sequence. Electrophoretic mobility shift assay (EMSA) showed that both clusters of zinc fingers bound to the same DNA sequence. Site-directed mutagenesis revealed that the core sequence TTGGTT within the GT box was essential to ZEC binding, while DNA sequences outside of the core sequence enhanced this interaction. Furthermore, co-transfection assays demonstrated that ZEC could activate a reporter luciferase gene driven by this DNA sequence.

Original languageEnglish (US)
Pages (from-to)71-81
Number of pages11
JournalGene
Volume340
Issue number1
DOIs
StatePublished - Sep 29 2004
Externally publishedYes

Fingerprint

Zinc Fingers
Proteins
Electrophoretic Mobility Shift Assay
Embryonic Stem Cells
Site-Directed Mutagenesis
Luciferases
Reporter Genes
Oligonucleotides
Transfection
Testis
Organism Cloning
Epitopes
Transcription Factors
Complementary DNA
Binding Sites
Kidney
Liver

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

ZEC, a zinc finger protein with novel binding specificity and transcription regulatory activity. / Chen, Guoyun; Muramatsu, Hisako; Ichihara-Tanaka, Keiko; Muramatsu, Takashi.

In: Gene, Vol. 340, No. 1, 29.09.2004, p. 71-81.

Research output: Contribution to journalArticle

Chen, Guoyun ; Muramatsu, Hisako ; Ichihara-Tanaka, Keiko ; Muramatsu, Takashi. / ZEC, a zinc finger protein with novel binding specificity and transcription regulatory activity. In: Gene. 2004 ; Vol. 340, No. 1. pp. 71-81.
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