Human disease-causing NOG missense mutations: effects on noggin secretion, dimer formation, and bone morphogenetic protein binding.

Secreted noggin protein regulates bone morphogenetic protein activity during development. In mice, a complete loss of noggin protein leads to multiple malformations including joint fusion, whereas mice heterozygous for Nog loss-of-function mutations are normal. In humans, heterozygous NOG missense mutations have been found in patients with two autosomal dominant disorders of joint development, multiple synostosis syndrome (SYNS1) and a milder disorder proximal symphalangism (SYM1). This study investigated the effect of one SYNS1 and two SYM1 disease-causing missense mutations on the structure and function of noggin. The SYNS1 mutation abolished, and the SYM1 mutations reduced, the secretion of functional noggin dimers in transiently transfected COS-7 cells. Coexpression of mutant noggin with wild-type noggin, to resemble the heterozygous state, did not interfere with wild-type noggin secretion. These data indicate that the human disease-causing mutations are hypomorphic alleles that reduce secretion of functional dimeric noggin. Therefore, we conclude that noggin has both species-specific and joint-specific dosage-dependent roles during joint formation. Surprisingly, in contrast to the COS-7 cell studies, the SYNS1 mutant was able to form dimers in Xenopus laevis oocytes. This finding indicates that there also exist species-specific differences in the ability to process mutant noggin polypeptides.

Relationship between defective mouse mammary tumor virus envelope glycoprotein synthesis and GRP78 synthesis in glucocorticoid-treated mouse lymphoma cells. Evidence for translational control of GRP78 synthesis.

We have found that synthesis of the 78-kDa glucose-regulated protein, GRP78, was increased following glucocorticoid treatment of S49 and W7MG1 mouse lymphoma cell lines. These lines synthesized a 74-kDa mouse mammary tumor virus envelope glycoprotein precursor, Pr74, both constitutively and in response to glucocorticoid treatment. In these cell lines, nascent Pr74 was not processed normally but was retained within the endoplasmic reticulum where it was bound stably by GRP78. GRP78 synthesis was not increased following glucocorticoid treatment of the W7.2 mouse lymphoma cell line, which does not synthesize Pr74, suggesting that the increase in GRP78 synthesis following glucocorticoid treatment of S49 and W7MG1 cells was secondary to the glucocorticoid-induced increase in Pr74 synthesis. Consistent with this hypothesis, the glucocorticoid-induced increase in Pr74 synthesis preceded the increase in GRP78 synthesis. Also, there was a direct correlation between the level of GRP78 synthesis and the level of Pr74 synthesis among multiple subclones of the W7-ENV/N line. This line was derived from the W7.2 line by retroviral mediated transfer of a constitutively expressed sequence encoding a defective form of Pr74 that was bound stably by GRP78 within the endoplasmic reticulum. An elevation in the steady state level of GRP78 mRNA was not detected either in glucocorticoid-treated S49 cells and W7MG1 cells or in subclones of the W7-ENV/N line that have an increased level of GRP78 synthesis. Therefore, it appears that the binding of Pr74 to GRP78 induces an increase in GRP78 synthesis that is regulated at a translational level, rather than at a transcriptional level.