NDLI: Mutations that affect the ability of the vnd/NK-2 homeoprotein to regulate gene expression: Transgenic alterations and tertiary structure

Content Provider	PubMed Central
Author	Koizumi, Keita Lintas, Carla Nirenberg, Marshall Maeng, Jin-soo Ju, Jeong-ho Mack, James W. Gruschus, James M. Odenwald, Ward F. Ferretti, James A.
Copyright Year	2003
Abstract	The importance in downstream target regulation of tertiary structure and DNA binding specificity of the protein encoded by the vnd/NK-2 homeobox gene is analyzed. The ectopic expression patterns of WT and four mutant vnd/NK-2 genes are analyzed together with expression of two downstream target genes, ind and msh, which are down-regulated by vnd/NK-2. Three mutants are deletions of conserved regions (i.e., tinman motif, acidic motif, and NK-2 box), and the fourth, Y54M vnd/NK-2, corresponds to a single amino acid residue replacement in the homeodomain. Of the four ectopically expressed mutant genes examined, only the Y54M mutation inactivates the ability of the vnd/NK-2 homeodomain protein to repress ind and msh. The acidic motif deletion mutant slightly reduced the ability of the protein to repress ind and msh. By contrast, both tinman and NK-2 box deletion mutants behaved as functional vnd/NK-2 genes in their ability to repress ind and msh. The NMR-determined tertiary structures of the Y54M vnd/NK-2 homeodomain, both free and bound to DNA, are compared with the WT analog. The only structural difference observed for the mutant homeodomain is in the complex with DNA and involved closer interaction of the methionine-54 with A2, rather than with C3 of the (−) strand of the DNA. This subtle change in the homeodomain–DNA complex resulted in modifications of binding affinities to DNA. These changes resulting from a single amino acid residue replacement constitute the molecular basis for the phenotypic alterations observed on ectopic expression of the Y54M vnd/NK-2 gene during embryogenesis.
Related Links	http://dx.doi.org/10.1073/pnas.0438043100
Ending Page	3124
Page Count	6
Starting Page	3119
File Format	PDF
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	6
Volume Number	100
Language	English
Publisher	The National Academy of Sciences
Publisher Date	2003-03-18
Access Restriction	Open
Rights Holder	The National Academy of Sciences
Subject Keyword	General Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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