Identification of Novel DNA Binding Targets and Regulatory Domains of a Murine Tinman Homeodomain Factor, nkx-2.5
Copyright policy )Identification of Novel DNA Binding Targets and Regulatory Domains of a Murine Tinman Homeodomain Factor, nkx-2.5
A murine cardiac-specific homeodomain gene named csx (Komuro, I., and Izumo. S. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 8145-8149) and nkx-2.5 (Lints, T. J., Parsons, L. M., Hartley, L., Lyons, I., and Harvey, R. P. (1993) Development 119, 419-431) was identified as a potential vertebrate homologue of Drosophila tinman, a mesoderm determination factor required for insect heart formation (Bodmer, R. (1993) Development 118, 719-729). Bacterial expression of the nkx-2.5 homeodomain allowed us to identify downstream DNA targets from a library of randomly generated oligonucleotides. High affinity nkx-2.5 DNA binding sites, 5'-TNNAGTG-3', represented novel binding sequences, whereas intermediate and weaker affinity sites, 5'-C(A/T)TTAATTN-3', contained the typical 5'-TAAT-3' core required by most homeodomain factors for DNA binding. We also observed that nkx-2.5 served as a modest transcription activator in transfection assays done in 10T1/2 fibroblasts with multimerized binding sites linked to a luciferase reporter gene. Functional dissection of nkx-2.5 revealed a COOH-terminal inhibitory domain composed mainly of clusters of alanines and prolines, which appeared to mask a potent activation domain composed of hydrophobic and highly charged amino acids.
- Baylor College of Medicine United States
Homeodomain Proteins, Transcriptional Activation, Binding Sites, Base Sequence, Molecular Sequence Data, DNA, Xenopus Proteins, Mice, Structure-Activity Relationship, Mutation, Homeobox Protein Nkx-2.5, Animals, Amino Acid Sequence, Transcription Factors
Homeodomain Proteins, Transcriptional Activation, Binding Sites, Base Sequence, Molecular Sequence Data, DNA, Xenopus Proteins, Mice, Structure-Activity Relationship, Mutation, Homeobox Protein Nkx-2.5, Animals, Amino Acid Sequence, Transcription Factors
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