Mutation analysis of TRPS1 gene including core promoter, 5'UTR, and 3'UTR regulatory sequences with insight into their organization.
The TRPS1 protein is a potent regulator of proliferation, differentiation, and apoptosis. The TRPS1 gene aberrations are strongly associated with rare trichorhinophalangeal syndrome (TRPS) development. We have conducted MLPA analysis to capture deletion within the crucial 8q24.1 chromosomal region in combination with mutation analysis of TRPS1 gene including core promoter, 5'UTR, and 3'UTR sequences in nine TRPS patients. Low complexity or extent of untranslated regulatory sequences avoided them from analysis in previous studies. Amplicon based next generation sequencing used in our study bridge over these technical limitations. Finally, we have made extended in silico analysis of TRPS1 gene regulatory sequences organization. Single contiguous deletion and an intragenic deletion intervening several exons were detected. Mutation analysis revealed five TRPS1 gene aberrations (two structural rearrangements, two nonsense mutations, and one missense substitution) reaching the overall detection rate of 78%. Several polymorphic variants were detected within the analysed regulatory sequences but without proposed pathogenic effect. In silico analysis suggested alternative promoter usage and diverse expression effectivity for different TRPS1 transcripts. Haploinsufficiency of TRPS1 gene was responsible for most of the TRPS phenotype. Structure of TRPS1 gene regulatory sequences is indicative of generally low single allele expression and its tight control.
Department of Anthropology and Human Genetics, Faculty of Science, Charles University in Prague, Vinicna 7, 128 43 Prague, Czech Republic. Electronic address: roman.solc@natur.cuni.cz.
Roman
R
Solc
Department of Anthropology and Human Genetics, Faculty of Science, Charles University in Prague, Vinicna 7, 128 43 Prague, Czech Republic. Electronic address: Mickey.007@seznam.cz.
Michaela
M
Klugerova
Department of Molecular Endocrinology, Institute of Endocrinology, Narodni 8, 116 94 Prague, Czech Republic. Electronic address: jvcelak@endo.cz.
Josef
J
Vcelak
Institute of Biology and Medical Genetics, 1st Faculty of Medicine and General University Hospital, Charles University in Prague, Albertov 4, 128 00 Prague, Czech Republic. Electronic address: baxova@vfn.cz.
Alice
A
Baxova
Department of Molecular Endocrinology, Institute of Endocrinology, Narodni 8, 116 94 Prague, Czech Republic; Genetic Department Olsanska, 1st and 3rd Faculty of Medicine, Charles University in Prague, Olsanska 2666/7, 130 00 Prague, Czech Republic. Electronic address: honza.kuklik@volny.cz.
Miloslav
M
Kuklik
Genetika Ostrava s.r.o., Korenskeho 1317/12, 702 00 Ostrava, Czech Republic. Electronic address: jan.vseticka55@gmail.com.
Jan
J
Vseticka
Department of Medical Genetics, University Hospital Brno, Cernopolni 212/9, 625 00 Brno, Czech Republic. Electronic address: rbeharka@fnbrno.cz.
Rastislav
R
Beharka
Institute of Biology and Medical Genetics, 1st Faculty of Medicine and General University Hospital, Charles University in Prague, Albertov 4, 128 00 Prague, Czech Republic. Electronic address: khirs@lf1.cuni.cz.
Katerina
K
Hirschfeldova
D020413:3' Untranslated Regions / Q000235:genetics*; D020121:5' Untranslated Regions / Q000235:genetics*; D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D005260:Female; D057895:Haploinsufficiency; D006801:Humans; D015826:Langer-Giedion Syndrome / Q000235:genetics; D008297:Male; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012097:Repressor Proteins; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D055815:Young Adult
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; C108387:TRPS1 protein, human; D014157:Transcription Factors
Alternative splicing; Chromosomal rearrangement; Mutation; NGS; Promoter