Abstract
A family of basic-helix-loop-helix (bHLH) nuclear factors play important roles in controlling cell growth and differentiation as critical regulatory components in transcription. Here we describe molecular characterization of mesoderm-specific bHLH protein, POD-1/Capsulin. Transactivation property of POD-1/Capsulin was analyzed by the Gal4 fusion system in six mammalian cell lines. The results indicated that an activation property was shown in HT1080 and HeLa cells, but a repression activity in HepG2 cells. Mapping analysis for the transactivation and repression activities revealed that the C-terminal domain of POD-1/Capsulin is essential for the transactivation and both the N-terminal and C-terminal domains are contributed to the repression activities. Furthermore, in order to identify possible interactants of the POD-1/Capsulin, we performed yeast two-hybrid screen in a human kidney cDNA library, and identified a class A bHLH protein, ITF-2 as potential heterodimeric partner of the bHLH protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Kadesch T: Consequences of heteromeric interactions among helixloop-helix proteins. Cell Growth Differ 4: 49–55, 1993
Murre C, McCaw PS, Vaessin H, Caudy M, Jan LY, Jan YN, Cabrera CV, Buskin JN, Hauschka SD, Lassar AB, et al: Interactions between heterologous helix-loop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell 58: 537–544, 1989
Davis RL, Cheng PF, Lassar AB, Weintraub H: The MyoD DNA binding domain contains a recognition code for muscle-specific gene activation. Cell. 60: 733–746, 1990
Henthorn P, Kiledjian M, Kadesch T: Two distinct transcription factors that bind the immunoglobulin enhancer microE5/kappa 2 motif. Science 247: 467–470, 1990
Hu JS, Olson EN, Kingston RE: HEB, a helix-loop-helix protein related to E2A and ITF2 that can modulate the DNA-binding ability of myogenic regulatory factors. Mol Cell Biol 12: 1031–1042, 1992
Lassar AB, Davis RL, Wright WE, Kadesch T, Murre C, Voronova A, Baltimore D, Weintraub H: Functional activity of myogenic HLH proteins requires hetero-oligomerization with E12/E47-like proteins in vivo. Cell 66: 305–315, 1991
Quaggin SE, Vanden Heuvel GB, Igarash P: POD-1, a mesoderm-specific basic-helix-loop-helix protein expressed in mesenchymal and glomerular epithelial cells in the developing kidney. Mech Dev 71: 37–48, 1998
Lu J, Richardson JA, Olson EN: Capsulin: a novel bHLH transcription factor expressed in epicardial progenitors and mesenchyme of visceral organs. Mech Dev 3: 23–32, 1998
Hidai H, Bardales R, Goodwin R, Quertermous T, Quertermous EE: Cloning of capsulin, a basic helix-loop-helix factor expressed in progenitor cells of the pericardium and the coronary arteries. Mech Dev 73: 33–43, 1998
Yoshida E, Aratani S, Itou H, Miyagishi M, Takiguchi M, Osumu T, Murakami K, Fukamizu A: Functional association between CBP and HNF4 in trans-activation. Biochem Biophys Res Commun 241: 664–669, 1997
Fields S, Song O: A novel genetic system to detect protein-protein interactions. Nature 340: 245–246, 1989
Chien CT, Bartel PL, Sternglanz R, Fields S: The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci USA 88: 9578–9582, 1991
Miller JH: Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1972
Blackwell TK, Weintraub H: Differences and similarities in DNAbinding preferences of MyoD and E2A protein complexes revealed by binding site selection. Science 250: 1104–1110, 1990
Poortinga G, Watanabe M, Parkhurst SM: Drosophila CtBP: A Hairy-interacting protein required for embryonic segmentation and hairy-mediated transcriptional repression. EMBO J 17: 2067–2078, 1998
Ayer DE, Lawrence QA, Eisenman RN: Mad-Max transcriptional repression is mediated by ternary complex formation with mammalian homologs of yeast repressor Sin3. Cell 80: 767–776, 1995
Rao G, Alland L, Guida P, Schreiber-Agus N, Chen K, Chin L, Rochelle JM, Seldin MF, Skoultchi AI, DePinho RA: Mouse Sin3A interacts with and can functionally substitute for the amino-terminal repression of the Myc antagonist Mxil. Oncogene 12: 1165–1172, 1996
Kasten MM, Ayer DE, Stillman DJ: SIN3-dependent transcriptional repression by interaction with the Mad1 DNA-binding protein. Mol Cell Biol 16: 4215–4221, 1996
Yuan W, Condorelli G, Caruso M, Felsani A, Giordano A: Human p300 protein is a coactivator for the transcription factor MyoD. J Biol Chem 271: 9009–9013, 1996
Sartorelli V, Huang J, Hamamori Y, Kedes L: Molecular mechanisms of myogenic coactivation by p300: direct interaction with the activation domain of MyoD and with the MADS box of MEF2C. Mol Cell Biol 17: 1010–1026, 1997
Puri PL, Avantaggiati ML, Balsano C, Sang N, Graessmann A, Giordano A, Levrero M: p300 is required for MyoD-dependent cell cycle arrest and muscle-specific gene transcription. EMBO J 16: 369–383, 1997
Eckner R, Yao TP, Oldread E, Livingston DM: Interaction and functional collaboration of p300/CBP and bHLH proteins in muscle and B-cell differentiation. Genes Dev 10: 2478–2490, 1996
Sato S, Roberts K, Gambino G, Cook A, Kouzarides T, Goding CR: CBP/p300 as a co-factor for the Microphthalmia transcription factor. Oncogene 14: 3083–3092, 1997
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miyagishi, M., Hatta, M., Ohshima, T. et al. Cell type-dependent transactivation or repression of mesoderm-restricted basic helix-loop-helix protein, POD-1/Capsulin. Mol Cell Biochem 205, 141–147 (2000). https://doi.org/10.1023/A:1007057611868
Issue Date:
DOI: https://doi.org/10.1023/A:1007057611868