EGAGenomicsILLUMINA, Illumina HiSeq 2500Otherhttps://ega-archive.org/studies/EGAS00001002272EGAEGA study EGAS00001002272Exome readsEGArestricted<h4>Purpose</h4>The purpose of this study was to identify mutations that cause non-syndromic male infertility using whole exome sequencing of family cases.<h4>Methods</h4>We recruited a consanguineous Turkish family comprising nine siblings with male triplets; two of the triplets were infertile as well as one younger infertile brother. Whole exome sequencing (WES) performed on two azoospermic brothers identified a mutation in the melanoma antigen family B4 (MAGEB4) gene which was confirmed via Sanger sequencing and then screened for on control groups and unrelated infertile subjects. The effect of the mutation on messenger RNA (mRNA) and protein levels was tested after in vitro cell transfection. Structural features of MAGEB4 were predicted throughout the conserved MAGE domain.<h4>Results</h4>The novel single-base substitution (c.1041A>T) in the X-linked MAGEB4 gene was identified as a no-stop mutation. The mutation is predicted to add 24 amino acids to the C-terminus of MAGEB4. Our functional studies were unable to detect any effect either on mRNA stability, intracellular localization of the protein, or the ability to homodimerize/heterodimerize with other MAGE proteins. We thus hypothesize that these additional amino acids may affect the proper protein interactions with MAGEB4 partners.<h4>Conclusion</h4>The whole exome analysis of a consanguineous Turkish family revealed MAGEB4 as a possible new X-linked cause of inherited male infertility. This study provides the first clue to the physiological function of a MAGE protein.A no-stop mutation in MAGEB4 is a possible cause of rare X-linked azoospermia and oligozoospermia in a consanguineous Turkish family.Okutman Ozlem O, Muller Jean J, Skory Valerie V, Garnier Jean Marie JM, Gaucherot Angeline A, Baert Yoni Y, Lamour Valérie V, Serdarogullari Munevver M, Gultomruk Meral M, Röpke Albrecht A, Kliesch Sabine S, Herbepin Viviana V, Aknin Isabelle I, Benkhalifa Moncef M, Teletin Marius M, Bakircioglu Emre E, Goossens Ellen E, Charlet-Berguerand Nicolas N, Bahceci Mustafa M, Tüttelmann Frank F, Viville STéphane SOligoasthenoteratozoospermia, Networks, 2810411E12Rik, azoospermia, Sperm Counts, Family Member, Kinship, Family Research, Count, Low Sperm, Map-6, Cryptozoospermia, Network, Low Sperm Count, Low Sperm Counts, mMage-b4, Mutations, Family Members, Azoospermia (finding), Hypospermatogeneses, STOP, Stable tubule-only polypeptide, Oligospermia, Cryptospermia, Mage-b4, Life Cycle, Family Life Cycle., Mtap6, 145-kDa STOP, Cryptozoospermias, rare (European definition), Low, Filiation, MAP-6, Sperm Count, Counts, Kinship Network, STOP145, Research, Absent sperm in semen, Oligozoospermia, Hypospermatogenesis, CN716893, Life Cycles, Oligoasthenoteratozoospermias, Cryptospermias, Families, spermatogenic failure, Kinship Networks, Family Life Cycles, Family, Relatives, CT3.6Materials, Kinship, Turkish, Substitution, Etiology, 1B1, Mutations, dmTAF[[II]]230, anon-EST:Posey9, intracellular localization, STOP, Treatment Effect, Method, Antigen, Life Cycle, Whole Transcriptome, Transcriptome Sequencing, Infertility, Non Polyadenylated, Missense, Male Infertility, Unrelated to Intervention, amino acids, Kinship Network, TFIID TAF250, cel, Moods, Basen, DMAGE, Non-Synonymous Mutation, procedures, Unable To, Sub-Fertility, "male infertility" EXACT [SNOMEDCT_2005_07_31:155924001], infertility, Decay, Adverse Event Unrelated to Intervention, HtsF, Add, ADD, dTAF[[II]]230, Polyadenylated, Exome, familial, TAF200, Test, Brother, Malignant Melanomas, Procedure, Dmel_CG9325, alpha-amino carboxylic acids, predicted, add, htsRC, Base2, CG43443, Base1, Substitution Mutation, RNA Decay, Confirmed, RNA Instability, Polyadenylated Messenger RNA, Acid, Complete Transcriptome, Non Polyadenylated mRNA, HTS, Hts, protein_coding_transcript, Amino acids, maleate, GLI3FL, Unable to do, INSDC_feature:gene, Methodological, Pdn, Non-Polyadenylated mRNA, Exon Non-Synonymous Mutation, Treatment Outcome, Taf250, Material, etiology, Whole Exome Sequencing, Unable to Do, Acids, Nepal Bhasa, Not at All, Poly(A) RNA, "male infertility NOS (finding)" EXACT [SNOMEDCT_2005_07_31:198018002], CT3.6, TAF230, ADD-87, whole exome, In Vitro as Topic, Effects, Male Sterility, Ovhts, treatment effect, HtsRC, protein-containing complex, Therapy Effect, Turkish Language, In Vitro Testing, melanoma (disease), New Lesion, Structural, Stable tubule-only polypeptide, Unrelated, Gene Products, Missense Variant, l(2)k14523, Ovhts-RC, mRNA Transcript, Technique, Sanger sequencing, In Vitro Tests, Brothers, Negation, In, Complete, Exome Sequencings, dTAF[[II]]250, STOP145, cell, Naevocarcinoma, messenger RNA, Structure, Sequencing, DETECT, Study, dTAF250, template RNA, Whole Exome, PERFORMED, Possible, Whole, Appearance, mRNA Transcript Degradation, constitutitional genetic, New, DmelCG43443, RNA Degradation, NEW, Perform, SINGLE, Hts-RC, RNA, 2810411E12Rik, Complete Exome Sequencings, Family Research, Not at All Able, Sub Fertility, Affects, Proteins, Messenger RNA, BG:DS00004.13, alpha-amino acids, Cell, Conclusion, dTAF230, Family Members, Exome Sequencing, In Vitro Test, male" EXACT [ICD9CM_2006:606], macromolecular stability of mRNA, native protein, Confirmatory, Poly(A)+ mRNA, Malignant Melanoma, chemical analysis, INSDC_feature:mRNA, TAF[[II]]250/230, mRNA Instability, 145-kDa STOP, Vitro Testing, Mutation, Bph, Taf[[II]]250, New Lesion Identification, Single Person, male sterility, mRNA, Sibling, Protein., missense mutation, Missense Mutation Abnormality, Negated, CN716893, Sister, male human body, No, Gene Proteins, Causation, Outcome of Therapy, Families, Sterility, In Vitro Technique, TUR, Solitary, nucleobases, Attention Deficit Hyperactivity Disorder, Polyadenylated mRNA, Instability, Relatives, hereditary, Alone, Networks, AU023367, Male, determination, Adverse Event Possibly Related to Intervention, Feature, Aminosaeure, mRNA Stability, Amino acid, Possibly Related to Intervention, protein, Techniques, melanoma, treatment outcome, (2Z)-but-2-enedioate, protein aggregate, Newari, Ability Question, Effect, male, Family Life Cycle, Group, CG10059, single organism cellular localization, Protein Altering Variant, Identification, Complete Exome Sequencing, Polyadenylated Messenger, Whole Transcriptome Sequencing, bases, localization within cell, Groups, genetic, Novel, Methodological Studies, Ability To, Family Life Cycles, l(2)01103, Exonic Non-Synonymous Mutation, Transfections, sterility, Nucleobase, Aminokarbonsaeure, unable, TAFII-250, male reproductive system infertility disorder, TAF250/230, results, mMage-b4, TAFII250, transfect, Melanoma, Non, Not, Mage-b4, Genetic Materials, Mtap6, Filiation, Genetic Material, Confirmation, cellular localisation, Possibly Related, Amino Acid, In Vitro, l(2)k06121, Tests, AI854843, conclusion, add-like, Control, Possible Attribution, mRNA Degradation, establishment and maintenance of cellular localization, Transcript Degradation, infertility disorder of male reproductive system, CG17603, Features, Methodological Study, TAF[[II]], Identified, Life Cycles, Adducin, "Infertility, Istanbul Turkish, Purpose, unspecified" EXACT [ICD9CM_2006:606.9], SR3-5, Base, Sisters, Cistron, inherited genetic, Male Sub-Fertility, Transcriptome Sequencings, base, protein levels, Non-, d230, Family Member, Procedures, Testings, establishment and maintenance of localization in cell or cell membrane, Complete Exome, Degradation, Aminocarbonsaeure, Gene, dTAFII250, CG9325, Network, alpha-amino acid, Malignant, EfW1, Xt, Subfertility, Stability, Missense Mutation, Therapy Outcome, dmTAF1, Messenger, Taf230, Add-hts, "male infertility (finding)" EXACT [SNOMEDCT_2005_07_31:2904007], Studies, l(2)00634, Mood, "male infertility, TAF250, study, WES, Taf200, HTS-R1, Genetic, single-organism cellular localization, Research, HTS-RC, Taf1p, DmelCG10059, dye terminator sequencing, Unrelated Attribution, Male Subfertility, Newar Language, male infertility, Control Group, Characteristics, Kinship Networks, TAF, Family, Dosing Days to Detection, treatment_outcome, CG34197, TAF[[II]]250, Males, protein complex, malignant, Map-6, l(3)84Ab, Cause, Cistrons, Testing, Melanomas, Triplet, Characteristic, GLI3-190, p230, Protein, In Vitro Testings, TFIID, Dmel_CG34197, techniques, Polyadenylated RNA, effect, mRNA Decay, MAP-6, Performed, purpose, Poly(A)+ RNA, Complete Transcriptome Sequencing, TAF[[II]]230, Poly(A) Tail, adducin, Non-Polyadenylated, TAF[II]250, Amino, Protein Gene Products, DmelCG17603, Only, male reproductive system infertility, malignant melanoma, Single, Additional, assay, EST D, methodology, TAF1Oligoasthenoteratozoospermia, Networks, 2810411E12Rik, azoospermia, Sperm Counts, Family Member, Kinship, Family Research, Count, Low Sperm, Map-6, Cryptozoospermia, Network, Low Sperm Count, Low Sperm Counts, mMage-b4, Mutations, Family Members, Azoospermia (finding), Hypospermatogeneses, STOP, Stable tubule-only polypeptide, Oligospermia, Cryptospermia, Mage-b4, Life Cycle, Family Life Cycle., Mtap6, 145-kDa STOP, Cryptozoospermias, rare (European definition), Low, Filiation, MAP-6, Sperm Count, Counts, Kinship Network, STOP145, Research, Absent sperm in semen, Oligozoospermia, Hypospermatogenesis, CN716893, Life Cycles, Oligoasthenoteratozoospermias, Cryptospermias, Families, spermatogenic failure, Kinship Networks, Family Life Cycles, Family, Relatives, CT3.6Networks, Materials, Kinship, AU023367, Male, determination, Aminosaeure, mRNA Stability, Amino acid, protein, 1B1, Mutations, dmTAF[[II]]230, anon-EST:Posey9, Techniques, intracellular localization, STOP, Method, Hominids, Life Cycle, Whole Transcriptome, Transcriptome Sequencing, Infertility, (2Z)-but-2-enedioate, protein aggregate, Non Polyadenylated, male, Family Life Cycle, Male Infertility, Group, single organism cellular localization, amino acids, Kinship Network, TFIID TAF250, cel, Moods, Complete Exome Sequencing, Polyadenylated Messenger, Whole Transcriptome Sequencing, bases, Basen, localization within cell, Groups, procedures, Sub-Fertility, genetic, "male infertility" EXACT [SNOMEDCT_2005_07_31:155924001], Methodological Studies, Hominin, infertility, Family Life Cycles, l(2)01103, Decay, Transfections, HtsF, Hominid, Add, ADD, sterility, dTAF[[II]]230, Polyadenylated, Nucleobase, Aminokarbonsaeure, Exome, familial, TAF200, Brother, Procedure, TAFII-250, male reproductive system infertility disorder, TAF250/230, Dmel_CG9325, results, alpha-amino carboxylic acids, predicted, mMage-b4, add, TAFII250, transfect, htsRC, Base2, CG43443, Base1, Mage-b4, Genetic Materials, Mtap6, Filiation, RNA Decay, RNA Instability, Genetic Material, cellular localisation, Polyadenylated Messenger RNA, Amino Acid, Acid, Complete Transcriptome, Non Polyadenylated mRNA, HTS, Hts, protein_coding_transcript, l(2)k06121, Amino acids, maleate, GLI3FL, AI854843, add-like, Control, INSDC_feature:gene, Methodological, mRNA Degradation, establishment and maintenance of cellular localization, Transcript Degradation, infertility disorder of male reproductive system, CG17603, Methodological Study, TAF[[II]], Pdn, Non-Polyadenylated mRNA, Life Cycles, Adducin, "Infertility, Taf250, unspecified" EXACT [ICD9CM_2006:606.9], Material, SR3-5, Base, Whole Exome Sequencing, Sisters, Cistron, Acids, inherited genetic, Male Sub-Fertility, Transcriptome Sequencings, Poly(A) RNA, "male infertility NOS (finding)" EXACT [SNOMEDCT_2005_07_31:198018002], CT3.6, TAF230, humans, base, protein levels, ADD-87, whole exome, d230, Family Member, Procedures, establishment and maintenance of localization in cell or cell membrane, Complete Exome, Male Sterility, Ovhts, Degradation, Aminocarbonsaeure, Gene, dTAFII250, HtsRC, CG9325, Network, alpha-amino acid, protein-containing complex, Hominini, EfW1, Xt, Apes, Subfertility, Stability, Stable tubule-only polypeptide, dmTAF1, Messenger, Taf230, Add-hts, Homo, "male infertility (finding)" EXACT [SNOMEDCT_2005_07_31:2904007], Studies, Gene Products, l(2)k14523, l(2)00634, Mood, Ovhts-RC, mRNA Transcript, "male infertility, Technique, Sanger sequencing, TAF250, study, Brothers, WES, Taf200, Complete, HTS-R1, Exome Sequencings, dTAF[[II]]250, Genetic, STOP145, single-organism cellular localization, Research, cell, messenger RNA, HTS-RC, Pongidae, Taf1p, Sequencing, dye terminator sequencing, Hominins, Study, dTAF250, template RNA, Male Subfertility, Whole Exome, male infertility, Whole, Control Group, mRNA Transcript Degradation, Kinship Networks, TAF, Family, constitutitional genetic, CG34197, DmelCG43443, RNA Degradation, Hts-RC, RNA, 2810411E12Rik, TAF[[II]]250, Complete Exome Sequencings, Family Research, Sub Fertility, Males, protein complex, Affects, Map-6, Proteins, Messenger RNA, l(3)84Ab, BG:DS00004.13, alpha-amino acids, Cistrons, Homininus, Cell, dTAF230, Family Members, Exome Sequencing, Triplet, male" EXACT [ICD9CM_2006:606], GLI3-190, macromolecular stability of mRNA, native protein, Poly(A)+ mRNA, p230, Protein, chemical analysis, INSDC_feature:mRNA, TAF[[II]]250/230, mRNA Instability, TFIID, 145-kDa STOP, Dmel_CG34197, techniques, Polyadenylated RNA, mRNA Decay, Bph, MAP-6, Taf[[II]]250, Poly(A)+ RNA, male sterility, Complete Transcriptome Sequencing, TAF[[II]]230, mRNA, Poly(A) Tail, Sibling, Protein., adducin, Non-Polyadenylated, TAF[II]250, Amino, CN716893, Sister, male human body, Protein Gene Products, Gene Proteins, DmelCG17603, Families, male reproductive system infertility, Sterility, nucleobases, Attention Deficit Hyperactivity Disorder, assay, Ape, Polyadenylated mRNA, EST D, Instability, Relatives, hereditary, methodology, TAF1falseA no-stop mutation in MAGEB4 is a possible cause of rare X-linked azoospermia and oligozoospermia in a consanguineous Turkish familyPurpose: The purpose of this study was to identify mutations that cause non-syndromic male infertility using whole exome sequencing of family cases.Methods: We recruited a consanguineous Turkish family comprising nine siblings with male triplets; two of the triplets were infertile as well as one younger infertile brother. Whole exome sequencing (WES) performed on two azoospermic brothers, identified a mutation in the MAGEB4 gene which was confirmed via Sanger sequencing and then screened for on control groups and unrelated infertile subjects. The effect of the mutation on mRNA and protein levels were tested after in vitro cell transfection. Structural features of MAGEB4 were predicted throughout the conserved MAGE domain.Results: The novel single base substitution (c.1041A>T) in the X-linked MAGEB4 gene was identified as a no-stop mutation. The mutation is predicted to add 24 amino acids to the C-terminus of MAGEB4. Our functional studies were unable to detect any effect either on mRNA stability, intracellular localization of the protein or the ability to homo / heterodimerize with other MAGE proteins. We thus hypothesize that these additional amino acids may affect the proper protein interactions with MAGEB4 partners. Conclusion: The whole exome analysis of a consanguineous Turkish family revealed MAGEB4 as a possible new X-linked cause of inherited male infertility. This study provides the first clue to the physiological function of a MAGE protein.2017-07-26 15:39:29EGAS00001002272960628401488EGAD00001003193EGAC00001000599