An 147L substitution in the HOXD13 homeodomain causes a novel human limb malformation by producing a selective loss of function

被引:58
作者
Caronia, G
Goodman, FR
McKeown, CME
Scambler, PJ
Zappavigna, V
机构
[1] DIBIT H San Raffaele, Dept Mol Biol & Funct Genom, I-20132 Milan, Italy
[2] Inst Child Hlth, Mol Med Unit, London WC1N 1EH, England
[3] Birmingham Maternity Hosp, Clin Genet Unit, Birmingham B15 2TG, W Midlands, England
[4] Univ Modena, Dept Anim Biol, I-41100 Modena, Italy
来源
DEVELOPMENT | 2003年 / 130卷 / 08期
关键词
Hox genes; limb malformations; missense mutation; DNA binding; posterior prevalence;
D O I
10.1242/dev.00396
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
The 5' members of the Hoxa and Hoxd gene clusters play major roles in vertebrate limb development. One such gene, HOXD]3, is mutated in the human limb malformation syndrome synpolydactyly. Both polyalanine tract expansions and frameshifting deletions in HOXD13 cause similar forms of this condition, but it remains unclear whether other kinds of HOXD13 mutations could produce different phenotypes. We describe a six-generation family in which a novel combination of brachydactyly and central polydactyly co-segregates with a missense mutation that substitutes leucine for isoleucine at position 47 of the HOXD13 homeodomain. We compared the HOXD13(I47L) mutant protein both in vitro and in vivo to the wild-type protein and to an artificial HOXD13 mutant, HOXD13(IQN), which is completely unable to bind DNA. We found that the mutation causes neither a dominant-negative effect nor a gain of function, but instead impairs DNA binding at some sites bound by wild-type HOXD13. Using retrovirus-mediated misexpression in developing chick limbs, we showed that wild-type HOXD13 could upregulate chick EphA7 in the autopod, but that HOXD13(I47L) could not. In the zeugopod, however, HOXD13(I47L) produced striking changes in tibial morphology and ectopic cartilages, which were never produced by HOXD13(IQN), consistent with a selective rather than generalised loss of function. Thus, a mutant HOX protein that recognises only a subset of sites recognised by the wild-type protein causes a novel human malformation, pointing to a hitherto undescribed mechanism by which missense mutations in transcription factors can generate unexpected phenotypes. Intriguingly, both HOXD13(I47L) and HOXD13(IQN) produced more severe shortening in proximal limb regions than did wildtype HOXD13, suggesting that functional suppression of anterior Hox genes by more posterior ones does not require DNA binding and is mediated by protein:protein interactions.
引用
收藏
页码:1701 / 1712
页数:12
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