Limits...
Unusual Stüve-Wiedemann syndrome with complete maternal chromosome 5 isodisomy.

Melone MA, Pellegrino MJ, Nolano M, Habecker BA, Johansson S, Nathanson NM, Knappskog PM, Hahn AF, Boman H - Ann Clin Transl Neurol (2014)

Bottom Line: A woman was isozygous for a novel mutation in the leukemia inhibitory factor receptor gene (LIFR) (c.2170C>G; p.Pro724Ala) which disrupts LIFR downstream signaling and results in Stüve-Wiedemann syndrome (STWS).She inherited two identical chromosomes 5 from her mother, heterozygous for the LIFR mutation.Prominent cold-induced sweating and heat intolerance lead to an initial diagnosis of cold-induced sweating syndrome, excluded by exome sequencing.

View Article: PubMed Central - PubMed

Affiliation: Division of Neurology and InterUniversity Center for Research in Neuroscience, Department of Clinical and Experimental Medicine and Surgery, Second University of Naples Naples, Italy.

ABSTRACT
A woman was isozygous for a novel mutation in the leukemia inhibitory factor receptor gene (LIFR) (c.2170C>G; p.Pro724Ala) which disrupts LIFR downstream signaling and results in Stüve-Wiedemann syndrome (STWS). She inherited two identical chromosomes 5 from her mother, heterozygous for the LIFR mutation. The presentation was typical for STWS, except there was no long bone dysplasia. Prominent cold-induced sweating and heat intolerance lead to an initial diagnosis of cold-induced sweating syndrome, excluded by exome sequencing. Skin biopsies provide the first human evidence of failed postnatal cholinergic differentiation of sympathetic neurons innervating sweat glands in cold-induced sweating, and of a neuropathy.

No MeSH data available.


Related in: MedlinePlus

Leukemia inhibitory factor receptor (LIFR) P724A mutant has altered glycosylation and impaired signaling. LIFR-negative Hep3B cells were transfected in duplicate with pcDNA3 (Vector, lanes 1–2), wild-type LIFR (wild-type, lanes 3–4), or the LIFR mutant (P724A, lanes 5–6), and blotted for LIFR. Molecular weight standards are marked on the left (kD). (A) Asterisks denote LIFR bands. The P724A mutant lacks the highest molecular weight band. (B) Cell lysates were combined with 5% SDS, 0.4 M DTT and denatured at 100°C for 10 min prior to incubation with 0.5 M sodium phosphate, pH 7.5, 1% NP40, and with (+) or without (−) PNGaseF (Peptide -N-Glycosidase F) for 1 hr at 37°C to remove glycosylation, and blotted for LIFR. Wild-type and P724A-transfected cells had LIFR of similar size after PNGaseF treatment, suggesting altered glycosylation of P724A-mutant receptor. (C) Transfected cells were treated with LIF to stimulate downstream signaling through LIFR-gp130 complex. Cells expressing wild-type LIFR exhibited strong STAT3 phosphorylation (Y705), but cells expressing vector alone or the P724A mutant did not. Total STAT3 (STAT3) was readily detectable in all cells. (D) LIFR protein was abundant in P724A transfected cells, despite the lack of STAT3 phosphorylation. All blots are representative of at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4265064&req=5

fig03: Leukemia inhibitory factor receptor (LIFR) P724A mutant has altered glycosylation and impaired signaling. LIFR-negative Hep3B cells were transfected in duplicate with pcDNA3 (Vector, lanes 1–2), wild-type LIFR (wild-type, lanes 3–4), or the LIFR mutant (P724A, lanes 5–6), and blotted for LIFR. Molecular weight standards are marked on the left (kD). (A) Asterisks denote LIFR bands. The P724A mutant lacks the highest molecular weight band. (B) Cell lysates were combined with 5% SDS, 0.4 M DTT and denatured at 100°C for 10 min prior to incubation with 0.5 M sodium phosphate, pH 7.5, 1% NP40, and with (+) or without (−) PNGaseF (Peptide -N-Glycosidase F) for 1 hr at 37°C to remove glycosylation, and blotted for LIFR. Wild-type and P724A-transfected cells had LIFR of similar size after PNGaseF treatment, suggesting altered glycosylation of P724A-mutant receptor. (C) Transfected cells were treated with LIF to stimulate downstream signaling through LIFR-gp130 complex. Cells expressing wild-type LIFR exhibited strong STAT3 phosphorylation (Y705), but cells expressing vector alone or the P724A mutant did not. Total STAT3 (STAT3) was readily detectable in all cells. (D) LIFR protein was abundant in P724A transfected cells, despite the lack of STAT3 phosphorylation. All blots are representative of at least three independent experiments.

Mentions: Transfection of wild-type (WT) LIFR into Hep3B cells generated LIFR protein of three different molecular weights, while cells transfected with the Pro724Ala mutant (LIFRP724A) contained only the smaller two forms of LIFR (Fig. 3A, asterisks). Glycosidase treatment with PNGaseF caused a downward shift in the higher molecular weight forms of both the WT and LIFR mutant to similar sizes (Fig. 3B), demonstrating that receptor glycosylation was altered in the Pro724Ala mutant. To determine if the mutant altered LIFR function, cells were treated with leukemia inhibitory factor (LIF) to stimulate downstream STAT3 phosphorylation. LIF stimulated robust STAT3 phosphorylation in WT LIFR-transfected cells (Fig. 3C), but little STAT3 phosphorylation in LIFRP724A transfected cells. The presence of STAT3 (Fig. 3C) and LIFR (Fig. 3D) protein was confirmed by western blot.


Unusual Stüve-Wiedemann syndrome with complete maternal chromosome 5 isodisomy.

Melone MA, Pellegrino MJ, Nolano M, Habecker BA, Johansson S, Nathanson NM, Knappskog PM, Hahn AF, Boman H - Ann Clin Transl Neurol (2014)

Leukemia inhibitory factor receptor (LIFR) P724A mutant has altered glycosylation and impaired signaling. LIFR-negative Hep3B cells were transfected in duplicate with pcDNA3 (Vector, lanes 1–2), wild-type LIFR (wild-type, lanes 3–4), or the LIFR mutant (P724A, lanes 5–6), and blotted for LIFR. Molecular weight standards are marked on the left (kD). (A) Asterisks denote LIFR bands. The P724A mutant lacks the highest molecular weight band. (B) Cell lysates were combined with 5% SDS, 0.4 M DTT and denatured at 100°C for 10 min prior to incubation with 0.5 M sodium phosphate, pH 7.5, 1% NP40, and with (+) or without (−) PNGaseF (Peptide -N-Glycosidase F) for 1 hr at 37°C to remove glycosylation, and blotted for LIFR. Wild-type and P724A-transfected cells had LIFR of similar size after PNGaseF treatment, suggesting altered glycosylation of P724A-mutant receptor. (C) Transfected cells were treated with LIF to stimulate downstream signaling through LIFR-gp130 complex. Cells expressing wild-type LIFR exhibited strong STAT3 phosphorylation (Y705), but cells expressing vector alone or the P724A mutant did not. Total STAT3 (STAT3) was readily detectable in all cells. (D) LIFR protein was abundant in P724A transfected cells, despite the lack of STAT3 phosphorylation. All blots are representative of at least three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4265064&req=5

fig03: Leukemia inhibitory factor receptor (LIFR) P724A mutant has altered glycosylation and impaired signaling. LIFR-negative Hep3B cells were transfected in duplicate with pcDNA3 (Vector, lanes 1–2), wild-type LIFR (wild-type, lanes 3–4), or the LIFR mutant (P724A, lanes 5–6), and blotted for LIFR. Molecular weight standards are marked on the left (kD). (A) Asterisks denote LIFR bands. The P724A mutant lacks the highest molecular weight band. (B) Cell lysates were combined with 5% SDS, 0.4 M DTT and denatured at 100°C for 10 min prior to incubation with 0.5 M sodium phosphate, pH 7.5, 1% NP40, and with (+) or without (−) PNGaseF (Peptide -N-Glycosidase F) for 1 hr at 37°C to remove glycosylation, and blotted for LIFR. Wild-type and P724A-transfected cells had LIFR of similar size after PNGaseF treatment, suggesting altered glycosylation of P724A-mutant receptor. (C) Transfected cells were treated with LIF to stimulate downstream signaling through LIFR-gp130 complex. Cells expressing wild-type LIFR exhibited strong STAT3 phosphorylation (Y705), but cells expressing vector alone or the P724A mutant did not. Total STAT3 (STAT3) was readily detectable in all cells. (D) LIFR protein was abundant in P724A transfected cells, despite the lack of STAT3 phosphorylation. All blots are representative of at least three independent experiments.
Mentions: Transfection of wild-type (WT) LIFR into Hep3B cells generated LIFR protein of three different molecular weights, while cells transfected with the Pro724Ala mutant (LIFRP724A) contained only the smaller two forms of LIFR (Fig. 3A, asterisks). Glycosidase treatment with PNGaseF caused a downward shift in the higher molecular weight forms of both the WT and LIFR mutant to similar sizes (Fig. 3B), demonstrating that receptor glycosylation was altered in the Pro724Ala mutant. To determine if the mutant altered LIFR function, cells were treated with leukemia inhibitory factor (LIF) to stimulate downstream STAT3 phosphorylation. LIF stimulated robust STAT3 phosphorylation in WT LIFR-transfected cells (Fig. 3C), but little STAT3 phosphorylation in LIFRP724A transfected cells. The presence of STAT3 (Fig. 3C) and LIFR (Fig. 3D) protein was confirmed by western blot.

Bottom Line: A woman was isozygous for a novel mutation in the leukemia inhibitory factor receptor gene (LIFR) (c.2170C>G; p.Pro724Ala) which disrupts LIFR downstream signaling and results in Stüve-Wiedemann syndrome (STWS).She inherited two identical chromosomes 5 from her mother, heterozygous for the LIFR mutation.Prominent cold-induced sweating and heat intolerance lead to an initial diagnosis of cold-induced sweating syndrome, excluded by exome sequencing.

View Article: PubMed Central - PubMed

Affiliation: Division of Neurology and InterUniversity Center for Research in Neuroscience, Department of Clinical and Experimental Medicine and Surgery, Second University of Naples Naples, Italy.

ABSTRACT
A woman was isozygous for a novel mutation in the leukemia inhibitory factor receptor gene (LIFR) (c.2170C>G; p.Pro724Ala) which disrupts LIFR downstream signaling and results in Stüve-Wiedemann syndrome (STWS). She inherited two identical chromosomes 5 from her mother, heterozygous for the LIFR mutation. The presentation was typical for STWS, except there was no long bone dysplasia. Prominent cold-induced sweating and heat intolerance lead to an initial diagnosis of cold-induced sweating syndrome, excluded by exome sequencing. Skin biopsies provide the first human evidence of failed postnatal cholinergic differentiation of sympathetic neurons innervating sweat glands in cold-induced sweating, and of a neuropathy.

No MeSH data available.


Related in: MedlinePlus