ABCMdb

PMID: 25010724

Sharma H, Mavuduru RS, Singh SK, Prasad R
Increased frequency of CFTR gene mutations identified in Indian infertile men with non-CBAVD obstructive azoospermia and spermatogenic failure.
Gene. 2014 Sep 10;548(1):43-7. doi: 10.1016/j.gene.2014.07.005. Epub 2014 Jul 7., [PubMed]

Sentences

No. Mutations Sentence Comment

5 ABCC7 p.Arg1358Ile ABCC7 p.Lys1351Arg Two mutations in exon 25 viz., R1358I and K1351R were identified as novel mutations in patients with non-CBAVD obstructive azoospermia. Login to comment 6 ABCC7 p.Arg1358Ile Mutation R1358I was predicted as probably damaging CFTR mutation. Login to comment 52 ABCC7 p.Arg117His ABCC7 p.Arg553* ABCC7 p.Asn1303Lys CFTR gene analysis, for the presence F508del, R117H, N1303K and R553X mutations was performed by single ARMS PCR as described by Ferrie et al. (1992). Login to comment 53 ABCC7 p.Gly551Asp ABCC7 p.Trp1282* ABCC7 p.Gly542* Other common mutations viz., 621+1GNT, G542X, G551D and W1282X were screened by multiplex ARMS PCR (Ferrie et al., 1992). Login to comment 87 ABCC7 p.Arg1358Ile ABCC7 p.Lys1351Arg SSCP analysis and subsequent DNA sequencing revealed two novel mutations viz., R1358I and K1351R (Table 2). Login to comment 88 ABCC7 p.Lys1351Arg Notably K1351R mutation was found in association with IVS8-5T allele in one patient. Login to comment 97 ABCC7 p.Arg1358Ile harvard.edu/pph2/) for R1358I was more than 0.05 (threshold for pathological mutation) and therefore categorized as deleterious mutation which may probably affect CFTR structure and functions (Table 4). Login to comment 98 ABCC7 p.Arg1358Ile Another algorithm mmb program (http://mmb.pcb.ub.edu/) also validates predictions done by PolyPhen-2 and revealed R1358I as pathological mutation. Login to comment 99 ABCC7 p.Arg1358Ile ABCC7 p.Arg1358Ile The output prediction score with iStable program (http://predictor.nchu.edu.tw/iStable/indexSeq.php) for R1358I was also more than 0.5 (threshold for stability), thus predicting that substitution of arginine to isoleucine at 1358 position may affect the structural stability of protein (Table 4). Login to comment 108 ABCC7 p.Arg1358Ile ABCC7 p.Lys1351Arg of alleles Non-CBAVD obstructive azoospermia (n = 60) T5 Reduction of oligo T tract to 5T at 1342-6 Aberrant splicing Intron 8 19 F508del Deletion of 3 bp (CTTor TTT) between 1652 and 1655 Deletion of phenylalanine at 508 Exon 11 7 R1358Ia G to A at 4073 Arginine to isoleucine at 1358 Exon 25 1 K1351Ra A to G at 4052 Lysine to arginine at 1351 Exon 25 1 Oligospermia/non-obstructive azoospermia (n = 150) T5 Reduction of oligo T tract to 5T at 1342-6 Aberrant splicing Intron 8 27 F508del Deletion of 3 bp (CTTor TTT) between 1652 and 1655 Deletion of phenylalanine at 508 Exon 10 11 a Indicates novel substitution mutations. Login to comment 137 ABCC7 p.Arg1358Ile ABCC7 p.Lys1351Arg Mutation Non-CBAVD obstructive azoospermia (OA) (n = 60) Spermatogenic failure (SF) (n = 150) Healthy control (ctrl) (n = 100) Chi square-test P valueÌe; OA vs ctrl SF vs ctrl OA vs SF 1 F508del 7 (11.6%) 11 (7.3%) 0 0.0008 0.008 NS 2 IVS(8)-5T homozygous 4 (6.6%) 7 (4.6%) 0 0.01 0.04 NS 3 IVS(8)-5T heterozygous 11 (18.3%) 13 (8.6%) 7 (7%) 0.00 NS 0.0001 4 R1358I 1 (1.6%) 0 0 NS - NS 5 K1351R 1 (1.6%) 0 0 NS - NS Ìe; P b 0.05 is considered as significant. Table 4 Pathological and stability prediction of novel substitution mutations identified in Indian non-CBAVD obstructive azoospermia patients. Login to comment 138 ABCC7 p.Arg1358Ile ABCC7 p.Lys1351Arg Pathological/structural prediction Novel mutants identified R1358I K1351R Polyphen score 1.00 0.03 Pathological prediction Probable damaging Benign iStable score 0.716 0.551 Stability prediction Decrease Unaffected Polyphen-2 (http://genetics.bwh.harvard.edu/pph2/) was used to generate the output score of novel sequence variants (threshold for pathological mutation was 0.05). Login to comment