Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Int J Mol Sci
2020 Mar 30;217:. doi: 10.3390/ijms21072382.
Show Gene links
Show Anatomy links
Novel Mutations in the TMPRSS3 Gene may Contribute to Taiwanese Patients with Nonsyndromic Hearing Loss.
Wong SH
,
Yen YC
,
Li SY
,
Yang JJ
.
???displayArticle.abstract???
A previous study indicated that mutations in the transmembrane protease serine 3 (TMPRSS3) gene, which encodes a transmembrane serine protease, cause nonsyndromic hearing loss (NSHL). This was the first description of a serine protease involved in hearing loss (HL). In Taiwan, however, data on the TMPRSS3 gene's association with NSHL is still insufficient. In this study, we described 10 mutations of TMPRSS3 genes found in 14 patients after screening 230 children with NSHL. The prevalence of the TMPRSS3 mutation appeared to be 6.09% (14/230). Of the 10 mutations, three were missense mutations: c.239G>A (p.R80H), c.551T>C (p.L184S), and 1253C>T (p.A418V); three were silent mutations, and four were mutations in introns. To determine the functional importance of TMPRSS3 mutations, we constructed plasmids carrying TMPRSS3 mutations of p.R80H, p.L184S, and p.A418V. TMPRSS3 function can be examined by secretory genetic assay for site-specific proteolysis (sGASP) and Xenopus oocyte expression system. Our results showed that p.R80H, p.L184S, and p.A418V TMPRSS3 mutations gave ratios of 19.4%, 13.2%, and 27.6%, respectively, via the sGASP system. Moreover, these three TMPRSS3 mutations failed to activate the epithelial sodium channel (ENaC) in the Xenopus oocyte expression system. These results indicate that the p.R80H, p.L184S, and p.A418V missense mutations of TMPRSS3 resulted in greatly diminishing the proteolytic activity of TMPRSS3. Our study provides information for understanding the importance of TMPRSS3 in the NSHL of Taiwanese children and provides a novel molecular explanation for the role of TMPRSS3 in HL.
Figure 1. Sequence analysis of TMPRSS3 genomic DNA variants in 230 patients with nonsyndromic hearing loss. Three variants of the TMPRSS3 gene were found in this study, three heterozygous missence variants: (A) c.239G>A, (B) c.551T>C, and (C) c.1253C>T. (D) Schematic representation of the domain structure of the TMPRSS3 protein with indication of the variants (black triangle). N: N-terminal domain; TM: transmembrane domains; LDLAR: low density lipoprotein receptor class A; SRCR: scavenger receptor cysteine rich; C: C-terminal domain.
Figure 2. Alignment of the amino acid sequences of TMPRSS3 proteins of various species. The (A) p.Arg80, (B) p.Leu184, and (C) p.Ala418 residues, which were variants in this study, are indicated in bold. Arg (R) at codon 80, Leu (L) at codon 184, and Ala (A) at codon 418 are identical among all species compared.
Figure 3. Assay of the catalytic activity of TMPRSS3 by site-specific proteolysis (sGASP). Transformants expressing both the STE13-substrate-invertase fusion protein and wild type (WT) or variants of the STE13-TMPRSS3 protein, including (A) TMPRSS3WT, (B) TMPRSS3R80H, (C) TMPRSS3L184S, (D) TMPRSS3A418V, and (E) TMPRSS3W251C, were plated on non-selective (glucose-YPD (yeast extract peptone dextrose)) and selective (sucrose-YPS (yeast extract peptone sucrose)) plates. TMPRSS3W251C mutant was as a positive control. (F) Quantitative assay of growth rate of yeast colony on sucrose plates. They were calculated by dividing the number of colony on sucrose plates by the number of colony on glucose plates (n = 5). ***: Asterisks denote the significant p-value <0.001.; STE13: Ste13p (Gene ID: 854394).
Figure 4. Assay of the Na+ currents of TMPRSS3 by electrophysiological measurements in Xenopus oocytes. The amiloride-sensitive current (INa) was measured in the presence of 120 μM of Na+ in Frog Ringer with 5 μM amiloride hydrochloride at a holding potential of 0 mV. (A) Due to the absence of any capped RNA (cRNA) as a negative control, the Na+ current was not produced and measured. (B) Co-injected cRNA of TMPRSS3WT and rat epithelial sodium channel (ENaC) α, β, and γ subunit into Xenopus oocytes, resulting in increased Na+ currents up to 5000–6000 nA (C) In contrast, the Na+ current was decreased to about 2000 nA in the amiloride hydrochloride treatment.
Figure 5. Functional expression of TMPRSS3 wild-type and mutant in Xenopus oocytes. Oocytes were injected with rat ENaC subunit in the presence of either (A) water (H2O), (B) TMPRSS3WT, and TMPRSS3 missense mutant, including (C) TMPRSS3R80H, (D) TMPRSS3L184S, (E) TMPRSS3A418V, and (F) TMPRSS3W251C. Water was used as a negative control. (G) All of the four missense mutant Na+ currents were similar to water, wherein there was no evident Na+ current and under 1000 nA in the quantitative analysis. In contrast, oocytes with the TMPRSS3WT emerging Na+ current were 5550 ± 1463 nA. (n ≥ 10).
Figure 6. Serine protease trypsin can activate ENaC to produce Na+ current in Xenopus oocytes. The three rat ENaC subunits and one of water, WT, or mutant TMPRSS3 were co-injected into Xenopus oocytes. After injection, the oocytes were treated with trypsin and then Na+ currents were recorded. (A) After trypsin treatment, the ENaC was significant activated, producing Na+ currents in the water (H2O) group. (B) In the TMPRSS3WT group, whether there was a trypsin treatment or not did not affect the ENaC activation and the Na+ currents. (C–F) All of the four missense mutation groups were very similar to the water group, and ENaC could be activated, increasing the Na+ currents. (G) In the quantitative analysis, 10 Xenopus oocytes were treated by trypsin and measured by ENaC-mediated INa in each group. (n ≥10).
Figure 7. Analysis of the effects of the TMPRSS3 mutant proteins on TMPRSS3WT by co-expression studies in the sGASP system and Xenopus oocytes. (A) Yeast (KSY01 strain) was transformed with substrate and equal amounts expression plasmids of TMPRSS3WT and TMPRSS3 mutants, as indicated and plated on YPD and YPS plates. Quantitative assay of growth rate was calculated by dividing the number of colony on YPS plates by the number of colony on YPD plates. All values are expressed as the mean ± SD of duplicate determinations (n = 5). (B) Rat ENaC subunit and equal amounts of TMPRSS3WT and TMPRSS3 mutant cRNA co-injected into Xenopus oocytes for electrophysiological ratios were calculated (n ≥ 10). These results indicated that the Na+ currents were under 2000 nA in the four heterozygous mutations group. Simultaneously, the ratios of Na+ currents were calculated by dividing the currents of TMPRSS3 mutant by the currents of TMPRSS3WT. All of the four heterozygous mutation ratios were less than 40% function of TMPRSS3WT. (C) After trypsin treatment, the ENaC was significant activated, producing Na+ currents in the heterozygous mutation type.
Apps,
Connexin 26 mutations in autosomal recessive deafness disorders: a review.
2007, Pubmed
Apps,
Connexin 26 mutations in autosomal recessive deafness disorders: a review.
2007,
Pubmed
Battelino,
TMPRSS3 mutations in autosomal recessive nonsyndromic hearing loss.
2016,
Pubmed
Ben-Yosef,
Novel mutations of TMPRSS3 in four DFNB8/B10 families segregating congenital autosomal recessive deafness.
2001,
Pubmed
Bonné-Tamir,
Linkage of congenital recessive deafness (gene DFNB10) to chromosome 21q22.3.
1996,
Pubmed
Bosanquet,
Acquired small bowel diverticular disease: a review.
2010,
Pubmed
Bugge,
Type II transmembrane serine proteases.
2009,
Pubmed
Canessa,
Amiloride-sensitive epithelial Na+ channel is made of three homologous subunits.
1994,
Pubmed
,
Xenbase
Cooper,
Where genotype is not predictive of phenotype: towards an understanding of the molecular basis of reduced penetrance in human inherited disease.
2013,
Pubmed
Donaudy,
Multiple mutations of MYO1A, a cochlear-expressed gene, in sensorineural hearing loss.
2003,
Pubmed
Fan,
Identification of a novel homozygous mutation, TMPRSS3: c.535G>A, in a Tibetan family with autosomal recessive non-syndromic hearing loss.
2014,
Pubmed
Gao,
Novel Mutations and Mutation Combinations of TMPRSS3 Cause Various Phenotypes in One Chinese Family with Autosomal Recessive Hearing Impairment.
2017,
Pubmed
Gao,
Identification of TMPRSS3 as a Significant Contributor to Autosomal Recessive Hearing Loss in the Chinese Population.
2017,
Pubmed
Gibson,
A type VII myosin encoded by the mouse deafness gene shaker-1.
1995,
Pubmed
Guipponi,
The transmembrane serine protease (TMPRSS3) mutated in deafness DFNB8/10 activates the epithelial sodium channel (ENaC) in vitro.
2002,
Pubmed
,
Xenbase
Guipponi,
TMPRSS3, a type II transmembrane serine protease mutated in non-syndromic autosomal recessive deafness.
2008,
Pubmed
Guipponi,
Mice deficient for the type II transmembrane serine protease, TMPRSS1/hepsin, exhibit profound hearing loss.
2007,
Pubmed
Guipponi,
An integrated genetic and functional analysis of the role of type II transmembrane serine proteases (TMPRSSs) in hearing loss.
2008,
Pubmed
Haerteis,
Proteolytic activation of the human epithelial sodium channel by trypsin IV and trypsin I involves distinct cleavage sites.
2014,
Pubmed
,
Xenbase
Hooper,
Type II transmembrane serine proteases. Insights into an emerging class of cell surface proteolytic enzymes.
2001,
Pubmed
Hutchin,
Assessment of the genetic causes of recessive childhood non-syndromic deafness in the UK - implications for genetic testing.
2005,
Pubmed
Jung,
Genetic Predisposition to Sporadic Congenital Hearing Loss in a Pediatric Population.
2017,
Pubmed
Kim,
Detection of site-specific proteolysis in secretory pathways.
2002,
Pubmed
Kubisch,
KCNQ4, a novel potassium channel expressed in sensory outer hair cells, is mutated in dominant deafness.
1999,
Pubmed
,
Xenbase
Lee,
Pathogenic mutations but not polymorphisms in congenital and childhood onset autosomal recessive deafness disrupt the proteolytic activity of TMPRSS3.
2003,
Pubmed
Lee,
Genetic analysis of TMPRSS3 gene in the Korean population with autosomal recessive nonsyndromic hearing loss.
2013,
Pubmed
Masmoudi,
Novel missense mutations of TMPRSS3 in two consanguineous Tunisian families with non-syndromic autosomal recessive deafness.
2001,
Pubmed
Morton,
Genetic epidemiology of hearing impairment.
1991,
Pubmed
Nesterov,
Trypsin can activate the epithelial sodium channel (ENaC) in microdissected mouse distal nephron.
2008,
Pubmed
Pallares-Ruiz,
A large deletion including most of GJB6 in recessive non syndromic deafness: a digenic effect?
2002,
Pubmed
Scott,
Insertion of beta-satellite repeats identifies a transmembrane protease causing both congenital and childhood onset autosomal recessive deafness.
2001,
Pubmed
Szabo,
Type II transmembrane serine proteases.
2003,
Pubmed
Veske,
Autosomal recessive non-syndromic deafness locus (DFNB8) maps on chromosome 21q22 in a large consanguineous kindred from Pakistan.
1996,
Pubmed
Wattenhofer,
Mutations in the TMPRSS3 gene are a rare cause of childhood nonsyndromic deafness in Caucasian patients.
2002,
Pubmed
Wattenhofer,
A novel TMPRSS3 missense mutation in a DFNB8/10 family prevents proteolytic activation of the protein.
2005,
Pubmed
,
Xenbase
Weegerink,
Genotype-phenotype correlation in DFNB8/10 families with TMPRSS3 mutations.
2011,
Pubmed
Weil,
Defective myosin VIIA gene responsible for Usher syndrome type 1B.
1995,
Pubmed
Wu,
Type II transmembrane serine proteases.
2003,
Pubmed
Yang,
Prospective variants screening of connexin genes in children with hearing impairment: genotype/phenotype correlation.
2010,
Pubmed
Yang,
Identification of mutations in members of the connexin gene family as a cause of nonsyndromic deafness in Taiwan.
2007,
Pubmed
Zlotogora,
Penetrance and expressivity in the molecular age.
2003,
Pubmed
de la Chapelle,
Genetic predisposition to human disease: allele-specific expression and low-penetrance regulatory loci.
2009,
Pubmed