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Enhancer of split^D, a dominant mutation of Drosophila, and its use in the study of functional domains of a helix-loop-helix protein

Tietze, Kyria and Oellers, Nadja and Knust, Elisabeth (1992) Enhancer of split^D, a dominant mutation of Drosophila, and its use in the study of functional domains of a helix-loop-helix protein. Proceedings of the National Academy of Sciences of the United States of America, 89 (13). pp. 6152-6156. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:20141216-144746126

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Abstract

Helix-loop-helix proteins play important roles in developmental processes, such as myogenesis, neurogenesis, and sex determination. The gene Enhancer of split [E(spl)] of Drosophila, a member of a gene complex that is involved in early neurogenesis, encodes a protein with a basic domain and a helix-loop-helix motif. We took advantage of a dominant mutation of this gene, E(spl)^D, to define in vivo structural features of this protein for proper function. The mutation renders the otherwise recessive eye phenotype of spl dominant. By germ-line transformation of different in vitro mutagenized versions of the E(spl) gene, we could demonstrate that the basic domain of this helix-loop-helix protein is functional and necessary for expression of the dominant phenotype. These results are supported by in vitro DNA-binding assays, which showed that the basic domain is in fact necessary for DNA binding, despite the presence of a proline residue. Furthermore, we could show that the dominant enhancement of spl is caused by truncation of the E(SPL)^D protein in combination with deletion of a putative regulatory element.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1073/pnas.89.13.6152DOIArticle
http://www.pnas.org/content/89/13/6152.abstractPublisherArticle
Additional Information:© 1992 National Academy of Sciences. Communicated by C. Nüsslein-Volhard, March 27, 1992. We are indebted to J. A. Campos-Ortega for many helpful discussions throughout the work and thank him and P. Hardy for constructive criticisms on the manuscript, F. Grawe for expert technical assistance, and F. W. Studier for the gift of pET expression vectors. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (SFB 243). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funders:
Funding AgencyGrant Number
Deutsche Forschungsgemeinschaft (DFG)SFB 243
Subject Keywords:DNA binding
Record Number:CaltechAUTHORS:20141216-144746126
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20141216-144746126
Official Citation: Enhancer of splitD, a dominant mutation of Drosophila, and its use in the study of functional domains of a helix-loop-helix protein K Tietze, N Oellers, and E Knust PNAS 1992 89 (13) 6152-6156
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52895
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:16 Dec 2014 23:03
Last Modified:16 Dec 2014 23:04

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