A zebrafish Ftz-F1 (Fushi Tarazu Factor 1) homologue requires multiple subdomains in the D and E regions for its transcriptional activity

Dong Liu, Mark Chandy, Soo-Kyung Lee, Yves Le Dréan, Hironori Ando, Fei Xiong, Jae Lee, Choy L. Hew

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

A zebrafish Ftz-F1 homologue, zFF1A (zebrafish Ff1a or Nr5a2, a member of nuclear receptor superfamily) and its C-terminally truncated variant (zFF1B) were previously identified. Due to lack of the identity box (I-box) and activation function 2 (AF-2) domain, zFF1B lacks transactivation function and fails to synergize with estrogen receptor (ER) in regulating promoters. It was speculated that the I-box might be involved in the zFF1A/ER interaction. In the present study, the function of the I-box was examined. In the absence of the I-box or with an altered heptad 9, the AF-2 of zFF1A was not functional, either in the presence or absence of ER. The GST pull-down assay showed that zFF1A and its mutants exerted similar physical contacts with ER-LBD, suggesting that the 'dimerization' domain (I-box) is essential for the transcriptional activity of zFF1A. Moreover, nuclear receptor coactivator selectively activated zFF1 with the I-box but exerted no effect on zFF1B, indicating that the I-box is able to interact with the coactivators. By deletion study and analysis of the identified domains in GAL4-DNA binding domain, other regions of zFF1A critical for its AF were also delineated. Consistent with the mutation analysis, AF-2 was active only in the presence of the I-box. We also identified a novel AF domain (AF-3) located in the hinge region (amino acids 155-267), although the activity of AF-3 was inhibited by its flanking region. We suggest that the D and E regions of zFF1A possess both positive and negative transactivation functions, and interdomain 'cross-talk' may confer the full transcriptional activity of the protein.

Original languageEnglish (US)
Pages (from-to)16758-16766
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number22
DOIs
StatePublished - Jun 2 2000
Externally publishedYes

Fingerprint

Steroidogenic Factor 1
E region
D region
Zebrafish
Estrogen Receptors
Transcriptional Activation
Nuclear Receptor Coactivators
Activation Analysis
Dimerization
Cytoplasmic and Nuclear Receptors
Chemical activation
Activation analysis
Hinges
Amino Acids
Mutation
Assays
DNA
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

A zebrafish Ftz-F1 (Fushi Tarazu Factor 1) homologue requires multiple subdomains in the D and E regions for its transcriptional activity. / Liu, Dong; Chandy, Mark; Lee, Soo-Kyung; Le Dréan, Yves; Ando, Hironori; Xiong, Fei; Lee, Jae; Hew, Choy L.

In: Journal of Biological Chemistry, Vol. 275, No. 22, 02.06.2000, p. 16758-16766.

Research output: Contribution to journalArticle

Liu, Dong ; Chandy, Mark ; Lee, Soo-Kyung ; Le Dréan, Yves ; Ando, Hironori ; Xiong, Fei ; Lee, Jae ; Hew, Choy L. / A zebrafish Ftz-F1 (Fushi Tarazu Factor 1) homologue requires multiple subdomains in the D and E regions for its transcriptional activity. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 22. pp. 16758-16766.
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abstract = "A zebrafish Ftz-F1 homologue, zFF1A (zebrafish Ff1a or Nr5a2, a member of nuclear receptor superfamily) and its C-terminally truncated variant (zFF1B) were previously identified. Due to lack of the identity box (I-box) and activation function 2 (AF-2) domain, zFF1B lacks transactivation function and fails to synergize with estrogen receptor (ER) in regulating promoters. It was speculated that the I-box might be involved in the zFF1A/ER interaction. In the present study, the function of the I-box was examined. In the absence of the I-box or with an altered heptad 9, the AF-2 of zFF1A was not functional, either in the presence or absence of ER. The GST pull-down assay showed that zFF1A and its mutants exerted similar physical contacts with ER-LBD, suggesting that the 'dimerization' domain (I-box) is essential for the transcriptional activity of zFF1A. Moreover, nuclear receptor coactivator selectively activated zFF1 with the I-box but exerted no effect on zFF1B, indicating that the I-box is able to interact with the coactivators. By deletion study and analysis of the identified domains in GAL4-DNA binding domain, other regions of zFF1A critical for its AF were also delineated. Consistent with the mutation analysis, AF-2 was active only in the presence of the I-box. We also identified a novel AF domain (AF-3) located in the hinge region (amino acids 155-267), although the activity of AF-3 was inhibited by its flanking region. We suggest that the D and E regions of zFF1A possess both positive and negative transactivation functions, and interdomain 'cross-talk' may confer the full transcriptional activity of the protein.",
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T1 - A zebrafish Ftz-F1 (Fushi Tarazu Factor 1) homologue requires multiple subdomains in the D and E regions for its transcriptional activity

AU - Liu, Dong

AU - Chandy, Mark

AU - Lee, Soo-Kyung

AU - Le Dréan, Yves

AU - Ando, Hironori

AU - Xiong, Fei

AU - Lee, Jae

AU - Hew, Choy L.

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