Molecular effects of novel mutations in Hesx1/HESX1 associated with human pituitary disorders
ABSTRACT The homeobox gene Hesx1/HESX1 has been implicated in the establishment of anterior pattern in the central nervous system (CNS) in a number of vertebrate species. Its role in pituitary development has been documented through loss-of-function studies in the mouse. A homozygous missense point mutation resulting in a single amino acid substitution, Arg160Cys (R160C), is associated with a heritable form of the human condition of septo-optic dysplasia (SOD). We have examined the phenotype of affected members in this pedigree in more detail and demonstrate for the first time a genetic basis for midline defects associated with an undescended or ectopic posterior pituitary. A similar structural pituitary abnormality was observed in a second patient heterozygous for another mutation in HESX1, Ser170Leu (S170L). Association of S170L with a pituitary phenotype may be a direct consequence of the HESX1 mutation since S170L is also associated with a dominant familial form of pituitary disease. However, a third mutation in HESX1, Asn125Ser (N125S), occurs at a high frequency in the Afro-Caribbean population and may therefore reflect a population-specific polymorphism. To investigate the molecular basis for these clinical phenotypes, we have examined the impact of these mutations on the regulatory functions of HESX1. We show that Hesx1 is a promoter-specific transcriptional repressor with a minimal 36 amino acid repression domain which can mediate promoter-specific repression by suppressing the activity of homeodomain-containing activator proteins. Mutations in HESX1 associated with pituitary disease appear to modulate the DNA-binding affinity of HESX1 rather than its transcriptional activity. Wild-type HESX1 binds a dimeric homeodomain site with high affinity (K(d) 31 nM) whilst HESX1(S170L) binds with a 5-fold lower activity (K(d) 150 nM) and HESX1(R160C) does not bind at all. Although HESX1(R160C) has only been shown to be associated with the SOD phenotype in children homozygous for the mutation, HESX1(R160C) can inhibit DNA binding by wild-type HESX1 both in vitro and in vivo in cell culture. This dominant negative activity of HESX1(R160C) is mediated by the Hesx1 repression domain, supporting the idea that the repression domain is implicated in interactions between homeodomain proteins. Our data suggest a possible molecular paradigm for the dominant inheritance observed in some pituitary disorders.
- SourceAvailable from: Alain Froment
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- "Additionally, the Pygmy 3p14.3 AIM haplotype includes a previously identified nonsynonymous SNP within HESX1 (rs9878928; Asn125Ser), which has been shown to be associated with pituitary developmental defects and growth hormone deficiency (Brickman et al., 2001; Gat-Yablonski et al., 2009). This nonsynonymous SNP is at moderate frequency (Pickrell et al., 2009) but on distinct haplotype backgrounds in other African populations . "
ABSTRACT: To reconstruct modern human evolutionary history and identify loci that have shaped hunter-gatherer adaptation, we sequenced the whole genomes of five individuals in each of three different hunter-gatherer populations at > 60× coverage: Pygmies from Cameroon and Khoesan-speaking Hadza and Sandawe from Tanzania. We identify 13.4 million variants, substantially increasing the set of known human variation. We found evidence of archaic introgression in all three populations, and the distribution of time to most recent common ancestors from these regions is similar to that observed for introgressed regions in Europeans. Additionally, we identify numerous loci that harbor signatures of local adaptation, including genes involved in immunity, metabolism, olfactory and taste perception, reproduction, and wound healing. Within the Pygmy population, we identify multiple highly differentiated loci that play a role in growth and anterior pituitary function and are associated with height.Cell 07/2012; 150(3):457-69. DOI:10.1016/j.cell.2012.07.009 · 33.12 Impact Factor
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- "The cDNAs used for ESC rescue were inserted into pCAGIP vector (Niwa et al., 2002; Niwa et al., 1991). Luciferase assays were performed as described previously (Brickman et al., 2001; Morrison and Brickman, 2006). ZHBTc4 ESCs (1 3 10 5 ) were plated on a 24-well plate with 2 mg/ml of Tc. "
ABSTRACT: Oct4 is an essential regulator of pluripotency in vivo and in vitro in embryonic stem cells, as well as a key mediator of the reprogramming of somatic cells into induced pluripotent stem cells. It is not known whether activation and/or repression of specific genes by Oct4 is relevant to these functions. Here, we show that fusion proteins containing the coding sequence of Oct4 or Xlpou91 (the Xenopus homolog of Oct4) fused to activating regions, but not those fused to repressing regions, behave as Oct4, suppressing differentiation and promoting maintenance of undifferentiated phenotypes in vivo and in vitro. An Oct4 activation domain fusion supported embryonic stem cell self-renewal in vitro at lower concentrations than that required for Oct4 while alleviating the ordinary requirement for the cytokine LIF. At still lower levels of the fusion, LIF dependence was restored. We conclude that the necessary and sufficient function of Oct4 in promoting pluripotency is to activate specific target genes.Cell Reports 02/2012; 1(2):99-109. DOI:10.1016/j.celrep.2011.12.002 · 7.21 Impact Factor
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- "Both homozygous and heterozygous missense mutations in HESX1 have been reported in association with SOD [Dattani et al., 1998; Thomas et al., 2001]. Mutations in HESX1 have also been reported in patients with ectopic posterior pituitary gland [Brickman et al., 2001] and in a patient with periventricular heterotopia and ectopic posterior pituitary gland [Mitchell et al., 2002]. The Hesx1 homeobox gene is expressed early in development in the mouse forebrain, and there is evidence for an important role for Hesx1/HESX1 in the development of mouse and human forebrain and pituitary gland [Dattani et al., 1998]. "
ABSTRACT: Schizencephaly is a malformation of cortical development characterized by gray matter-lined clefts in the cerebral cortex and a range of neurological presentations. In some cases, there are features of septo-optic dysplasia concurrently with schizencephaly. The etiologies of both schizencephaly and septo-optic dysplasia are thought to be heterogeneous, but there is evidence that at least some cases have genetic origin. We hypothesized that these disorders may be caused by mutations in three candidate genes: LHX2, a gene with an important cortical patterning role, and HESX1 and SOX2, genes that have been associated with septo-optic dysplasia. We sequenced a large cohort of patients with schizencephaly, some with features of septo-optic dysplasia, for mutations in these genes. No pathogenic mutations were observed, suggesting that other genes or non-genetic factors influencing genes critical to brain development must be responsible for schizencephaly.American Journal of Medical Genetics Part A 11/2010; 152A(11):2736-42. DOI:10.1002/ajmg.a.33684 · 2.05 Impact Factor