ABSTRACT
OBJECTIVE: To investigate the clinical and genetic features of a child with Dyschromatosis symmetrica hereditaria (DSH) and variant of the ADAR1 gene. METHODS: A child who was admitted to the Department of Dermatology of the First Affiliated Hospital of Zhengzhou University in June 2020 due to irregular pigmented maculopapular rash on the dorsum of hands was selected as the study subject. Whole exome sequencing (WES) was carried out for the child and his similarly affected father, and Sanger sequencing was used to verify the candidate variant. SWISS-MODEL was used to predict the secondary and tertiary structures of the wild-type and mutant ADAR1 proteins. RESULTS: The child, a 13-year-old boy, had symmetrical hyperpigmented and depigmented spots on the back of his hands and was clinically diagnosed with DSH. WES and Sanger sequencing results showed that he and his father had both harbored a heterozygous c.2858dup (p.T954Dfs*20) truncating variant in exon 10 of the ADAR1 gene. Based on the guidelines from the American College of Medical Genetics and Genomics, the variant was predicted as pathogenic (PVS1+PM2_Supporting+PM1+PP3). CONCLUSION: The c.2858dup (p.T954Dfs*20) variant of the ADAR1 gene probably underlay the DSH in this pedigree.
Subject(s)
Adenosine Deaminase , Pigmentation Disorders , RNA-Binding Proteins , Humans , Male , Adenosine Deaminase/genetics , Pigmentation Disorders/genetics , Pigmentation Disorders/congenital , RNA-Binding Proteins/genetics , Adolescent , Mutation , Exome Sequencing , Exons , Genetic Testing , PedigreeABSTRACT
The aim of this study was to compare the circular transcriptome of divergent tissues in order to understand: i) the presence of circular RNAs (circRNAs) that are not exonic circRNAs, i.e. originated from backsplicing involving known exons and, ii) the origin of artificial circRNA (artif_circRNA), i.e. circRNA not generated in-vivo. CircRNA identification is mostly an in-silico process, and the analysis of data from the BovReg project (https://www.bovreg.eu/) provided an opportunity to explore new ways to identify reliable circRNAs. By considering 117 tissue samples, we characterized 23,926 exonic circRNAs, 337 circRNAs from 273 introns (191 ciRNAs, 146 intron circles), 108 circRNAs from small non-coding genes and nearly 36.6K circRNAs classified as other_circRNAs. Furthermore, for 63 of those samples we analysed in parallel data from total-RNAseq (ribosomal RNAs depleted prior to library preparation) with paired mRNAseq (library prepared with poly(A)-selected RNAs). The high number of circRNAs detected in mRNAseq, and the significant number of novel circRNAs, mainly other_circRNAs, led us to consider all circRNAs detected in mRNAseq as artificial. This study provided evidence of 189 false entries in the list of exonic circRNAs: 103 artif_circRNAs identified by total RNAseq/mRNAseq comparison using two circRNA tools, 26 probable artif_circRNAs, and 65 identified by deep annotation analysis. Extensive benchmarking was performed (including analyses with CIRI2 and CIRCexplorer-2) and confirmed 94% of the 23,737 reliable exonic circRNAs. Moreover, this study demonstrates the effectiveness of a panel of highly expressed exonic circRNAs (5-8%) in analysing the tissue specificity of the bovine circular transcriptome.
Subject(s)
Exons , RNA, Circular , RNA, Circular/genetics , Animals , Cattle , Introns , Computational Biology/methods , Transcriptome , Gene Expression Profiling/methods , Sequence Analysis, RNA/methodsABSTRACT
The actin cytoskeleton is one of the most important players in cell motility, adhesion, division, and functioning. The regulation of specific microfilament formation largely determines cellular functions. The main actin-binding protein in animal cells is tropomyosin (Tpm). The unique structural and functional diversity of microfilaments is achieved through the diversity of Tpm isoforms. In our work, we studied the properties of the cytoplasmic isoforms Tpm1.8 and Tpm1.9. The results showed that these isoforms are highly thermostable and differ in the stability of their central and C-terminal fragments. The properties of these isoforms were largely determined by the 6th exons. Thus, the strength of the end-to-end interactions, as well as the affinity of the Tpm molecule for F-actin, differed between the Tpm1.8 and Tpm1.9 isoforms. They were determined by whether an alternative internal exon, 6a or 6b, was included in the Tpm isoform structure. The strong interactions of the Tpm1.8 and Tpm1.9 isoforms with F-actin led to the formation of rigid actin filaments, the stiffness of which was measured using an optical trap. It is quite possible that the structural and functional features of the Tpm isoforms largely determine the appearance of these isoforms in the rigid actin structures of the cell cortex.
Subject(s)
Actin Cytoskeleton , Actins , Protein Isoforms , Tropomyosin , Tropomyosin/metabolism , Tropomyosin/chemistry , Tropomyosin/genetics , Protein Isoforms/metabolism , Protein Isoforms/chemistry , Protein Isoforms/genetics , Actin Cytoskeleton/metabolism , Animals , Actins/metabolism , Actins/chemistry , Cytoplasm/metabolism , Humans , Exons , Protein Binding , Protein StabilityABSTRACT
Cefaclor is a substrate of human-peptide-transporter-1 (PEPT1), and the impact of inter-individual pharmacokinetic variation due to genetic polymorphisms of solute-carrier-family-15-member-1 (SLC15A1) has been a topic of great debate. The main objective of this study was to analyze and interpret cefaclor pharmacokinetic variations according to genetic polymorphisms in SLC15A1 exons 5 and 16. The previous cefaclor bioequivalence results were integrated with additional SLC15A1 exons 5 and 16 genotyping results. An analysis of the structure-based functional impact of SLC15A1 exons 5 and 16 genetic polymorphisms was recently performed using a PEPT1 molecular modeling approach. In cefaclor pharmacokinetic analysis results according to SLC15A1 exons 5 and 16 genetic polymorphisms, no significant differences were identified between genotype groups. Furthermore, in the population pharmacokinetic modeling, genetic polymorphisms in SLC15A1 exons 5 and 16 were not established as effective covariates. PEPT1 molecular modeling results also confirmed that SLC15A1 exons 5 and 16 genetic polymorphisms did not have a significant effect on substrate interaction with cefaclor and did not have a major effect in terms of structural stability. This was determined by comprehensively considering the insignificant change in energy values related to cefaclor docking due to point mutations in SLC15A1 exons 5 and 16, the structural change in conformations confirmed to be less than 0.05 Å, and the relative stabilization of molecular dynamic simulation energy values. As a result, molecular structure-based analysis recently suggested that SLC15A1 exons 5 and 16 genetic polymorphisms of PEPT1 were limited to being the main focus in interpreting the pharmacokinetic diversity of cefaclor.
Subject(s)
Cefaclor , Peptide Transporter 1 , Humans , Peptide Transporter 1/genetics , Peptide Transporter 1/metabolism , Cefaclor/pharmacokinetics , Exons/genetics , Genotype , Polymorphism, Genetic , Anti-Bacterial Agents/pharmacokinetics , Polymorphism, Single Nucleotide , Models, MolecularABSTRACT
Duchenne muscular dystrophy (DMD) is a severe X-linked recessive genetic disorder caused by mutations in the DMD gene, which leads to a deficiency of the dystrophin protein. The main mutation types of this gene include exon deletions and duplications, point mutations, and insertions. These mutations disrupt the normal expression of dystrophin, ultimately leading to the disease. In this study, we reported a case of DMD caused by an insertion mutation in exon 59 (E59) of the DMD gene. The affected child exhibited significant abnormalities in related biochemical markers, early symptoms of DMD, and multiple gray hair. His mother and sister were carriers with slightly abnormal biochemical markers. The mother had mild clinical symptoms, while the sister had no clinical symptoms. Other family members were genetically and physically normal. Sequencing and sequence alignment revealed that the inserted fragment was an Alu element from the AluYa5 subfamily. This insertion produced two stop codons and a polyadenylate (polyA) tail. To understand the impact of this insertion on the DMD gene and its association with clinical symptoms, exonic splicing enhancer (ESE) prediction indicated that the insertion did not affect the splicing of E59. Therefore, we speculated that the insertion sequence would be present in the mRNA sequence of the DMD gene. The two stop codons and polyA tail likely terminate translation, preventing the production of functional dystrophin protein, which may be the mechanism leading to DMD. In addition to typical DMD symptoms, the child also exhibited premature graying of hair. This study reports, for the first time, a case of DMD caused by the insertion of an Alu element into the coding region of the DMD gene. This finding provides clues for studying gene mutations induced by Alu sequence insertion and expands the understanding of DMD gene mutations.
Subject(s)
Alu Elements , Dystrophin , Muscular Dystrophy, Duchenne , Mutagenesis, Insertional , Muscular Dystrophy, Duchenne/genetics , Humans , Alu Elements/genetics , Dystrophin/genetics , Male , Base Sequence , Hair/metabolism , Female , Exons/genetics , Child , Molecular Sequence DataABSTRACT
The newly discovered HLA-C*04:01:01:186 allele differs from HLA-C*04:01:01:01 by a single nucleotide substitution in intron 3.
Subject(s)
Alleles , HLA-C Antigens , High-Throughput Nucleotide Sequencing , Histocompatibility Testing , Introns , Humans , HLA-C Antigens/genetics , High-Throughput Nucleotide Sequencing/methods , Histocompatibility Testing/methods , Greece , Polymorphism, Single Nucleotide , Exons , Base Sequence , Sequence Analysis, DNA/methodsABSTRACT
BACKGROUND: Synonymous variants are non-pathogenic due to non-substitution of amino acids. However, synonymous exonic terminal nucleotide substitutions may affect splicing. Splicing variants are easily analyzed at RNA level for genes expressed in blood cells. Minigene analysis provides another method for splicing variant analysis of genes that are poorly or not expressed in peripheral blood. METHODS: Whole exome sequencing was performed to screen for potential pathogenic mutations in the proband, which were validated within the family by Sanger sequencing. The pathogenicity of the synonymous mutation was analyzed using the minigene technology. RESULTS: The proband harbored the compound heterogeneous variants c. [291G >A; 572-50C >T] and c.681 + 1G >T in F7, of which the synonymous variant c.291G >A was located at the terminal position of exon 3. Minigene analysis revealed exon3 skipping due to this mutation, which may have subsequently affected protein sequence, structure, and function. CONCLUSION: Our finding confirmed the pathogenicity of c.291G >A, thus extending the pathogenic mutation spectrum of F7, and providing insights for effective reproductive counseling.
Subject(s)
Exons , RNA Splicing , Silent Mutation , Humans , Female , Male , Pedigree , AdultABSTRACT
We report the case of an 87-year-old female patient who was diagnosed with metastatic non-small-cell lung cancer harboring MET exon 14 skipping mutation (MET ex14) and PD-L1 expression of 60%. A first-line treatment with atezolizumab was started with primary resistance. Then, a second-line treatment with capmatinib, a selective type Ib MET tyrosine kinase inhibitor, was started, achieving a partial response. The patient is still alive and on treatment with capmatinib 300 mg twice daily after 20 months, with a good tolerability and no evidence of disease progression.In summary, our patient experienced a long-lasting response (>18 months) with capmatinib as second-line treatment. Further analyses evaluating the efficacy and tolerability of MET tyrosine kinase inhibitors are warranted, especially in the elderly, a non-small-cell lung cancer population whose tumors could more frequently harbor MET ex14 mutation.
[Box: see text].
Subject(s)
Carcinoma, Non-Small-Cell Lung , Exons , Lung Neoplasms , Mutation , Proto-Oncogene Proteins c-met , Humans , Female , Proto-Oncogene Proteins c-met/genetics , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Aged, 80 and over , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Mutation/genetics , Exons/genetics , Protein Kinase Inhibitors/therapeutic use , Protein Kinase Inhibitors/adverse effects , Protein Kinase Inhibitors/administration & dosage , Benzamides/therapeutic use , Benzamides/adverse effects , Treatment Outcome , Acrylamides/therapeutic use , Acrylamides/administration & dosage , Acrylamides/adverse effects , Antineoplastic Agents/therapeutic use , Antineoplastic Agents/adverse effects , Imidazoles , TriazinesABSTRACT
HLA-DPA1*02:134, a novel HLA class II allele detected by next-generation sequencing.
Subject(s)
Alleles , Exons , HLA-DP alpha-Chains , High-Throughput Nucleotide Sequencing , Histocompatibility Testing , Humans , High-Throughput Nucleotide Sequencing/methods , HLA-DP alpha-Chains/genetics , Histocompatibility Testing/methods , CodonABSTRACT
HLA-DQA1*05:03:03, a novel HLA class II allele detected by next-generation sequencing.
Subject(s)
Alleles , Exons , HLA-DQ alpha-Chains , High-Throughput Nucleotide Sequencing , Histocompatibility Testing , Humans , HLA-DQ alpha-Chains/genetics , High-Throughput Nucleotide Sequencing/methods , Histocompatibility Testing/methods , Codon , Base Sequence , Sequence Analysis, DNA/methodsABSTRACT
We describe the novel KIR3DL1*182 allele and confirmed the 3DL1*15002 allele.
Subject(s)
Alleles , High-Throughput Nucleotide Sequencing , Receptors, KIR3DL1 , Humans , Receptors, KIR3DL1/genetics , High-Throughput Nucleotide Sequencing/methods , Exons , Base SequenceABSTRACT
Duchenne muscular dystrophy (DMD) affecting 1 in 3500-5000 live male newborns is the frequently fatal genetic disease resulted from various mutations in DMD gene encoding dystrophin protein. About 70% of DMD-causing mutations are exon deletion leading to frameshift of open reading frame and dystrophin deficiency. To facilitate translating human DMD-targeting CRISPR therapeutics into patients, we herein establish a genetically humanized mouse model of DMD by replacing exon 50 and 51 of mouse Dmd gene with human exon 50 sequence. This humanized mouse model recapitulats patient's DMD phenotypes of dystrophin deficiency and muscle dysfunction. Furthermore, we target splicing sites in human exon 50 with adenine base editor to induce exon skipping and robustly restored dystrophin expression in heart, tibialis anterior and diaphragm muscles. Importantly, systemic delivery of base editor via adeno-associated virus in the humanized male mouse model improves the muscle function of DMD mice to the similar level of wildtype ones, indicating the therapeutic efficacy of base editing strategy in treating most of DMD types with exon deletion or point mutations via exon-skipping induction.
Subject(s)
Adenine , CRISPR-Cas Systems , Disease Models, Animal , Dystrophin , Exons , Gene Editing , Muscular Dystrophy, Duchenne , Animals , Muscular Dystrophy, Duchenne/genetics , Muscular Dystrophy, Duchenne/therapy , Dystrophin/genetics , Dystrophin/metabolism , Exons/genetics , Humans , Male , Gene Editing/methods , Mice , Adenine/metabolism , Muscle, Skeletal/metabolism , Dependovirus/genetics , Genetic Therapy/methodsABSTRACT
We talk to corresponding author Thomas Gonatopoulos-Pournatzis and co-first authors Arun Prasath Damodaran and Mei-Sheng Xiao about their paper "Genome-scale exon perturbation screens uncover exons critical for cell fitness" (in this issue of Molecular Cell) and get insights into their findings, career trajectories, and future directions in the pre-mRNA processing field.
Subject(s)
Exons , Humans , History, 21st Century , History, 20th Century , RNA Precursors/genetics , RNA Precursors/metabolism , RNA SplicingABSTRACT
HLA-C*04:01:01:182 differs from the HLA-C*04:01:01:06 allele by one nucleotide substitution in the 5'UTR.
Subject(s)
Alleles , HLA-C Antigens , High-Throughput Nucleotide Sequencing , Histocompatibility Testing , Tissue Donors , Humans , HLA-C Antigens/genetics , High-Throughput Nucleotide Sequencing/methods , Histocompatibility Testing/methods , 5' Untranslated Regions , Exons , Base Sequence , Sequence Analysis, DNA/methods , Bone Marrow , Polymorphism, Single Nucleotide , Bone Marrow TransplantationABSTRACT
HLA-DRB1*09:54 shows a substitution G to A at position 449 when compared with HLA-DRB1*09:01:02:01.
Subject(s)
Alleles , Asian People , Exons , HLA-DRB1 Chains , Histocompatibility Testing , Humans , HLA-DRB1 Chains/genetics , Asian People/genetics , Base Sequence , Sequence Analysis, DNA/methods , Amino Acid Substitution , Sequence Alignment , Polymorphism, Single Nucleotide , Codon , East Asian PeopleABSTRACT
Characterisation of the novel HLA-C*14:02:01:31 allele in a 21-year-old Greek bone marrow donor.
Subject(s)
Alleles , Exons , HLA-C Antigens , High-Throughput Nucleotide Sequencing , Histocompatibility Testing , Tissue Donors , Humans , HLA-C Antigens/genetics , Young Adult , Bone Marrow Transplantation , Bone Marrow , Base Sequence , Polymorphism, Single Nucleotide , CodonABSTRACT
One nucleotide substitution in codon 320 of A*03:01:01:01 results in the novel allele, HLA-A*03:478.
Subject(s)
Alleles , Asian People , Exons , Histocompatibility Testing , Humans , Asian People/genetics , Republic of Korea , Sequence Analysis, DNA/methods , Base Sequence , Codon , HLA-A3 Antigen/genetics , HLA-A Antigens/genetics , Polymorphism, Single Nucleotide , Sequence AlignmentABSTRACT
The novel HLA-C*01:273 allele, first described in a potential bone marrow donor from Brazil.
Subject(s)
Alleles , Exons , HLA-C Antigens , Humans , HLA-C Antigens/genetics , Brazil , Histocompatibility Testing , Base Sequence , Tissue Donors , Sequence Analysis, DNA/methods , Sequence Alignment , CodonABSTRACT
The novel HLA-A*30:221 allele differs from HLA-A*30:01:01:01 by one nucleotide substitution in Exon 7.