ABCMdb

PMID: 22337229

Agrawal A, Hamvas A, Cole FS, Wambach JA, Wegner D, Coghill C, Harrison K, Nogee LM
An intronic ABCA3 mutation that is responsible for respiratory disease.
Pediatr Res. 2012 Jun;71(6):633-7. doi: 10.1038/pr.2012.21. Epub 2012 Feb 15., [PubMed]

Sentences

No. Mutations Sentence Comment

30 ABCA3 p.Glu690Lys To determine whether the aberrantly spliced transcripts were derived from the allele with the c.2068 G>A (p.E690K) mutation, primers were designed to amplify either the transcripts containing the intronic sequence or those that were normally spliced, by using a 3' primer located in the intronic sequence, or one that spanned exons 25 and 26. Login to comment 33 ABCA3 p.Glu690Lys Electrophoretic analysis of the restriction products demonstrated that the relevant Ban II restriction fragment derived from the aberrantly spliced transcripts was smaller than that derived from the normally spliced products, indicating the presence of the G allele (wild type) in those amplicons containing the intronic sequences, and the A allele (p.E690K mutation) in those containing normal splicing. Login to comment 42 ABCA3 p.Glu690Lys The mother was found to be heterozygous GA at position c.2068 (site of the p.E690K mutation) but was homozygous CC at position IVS25-98. Login to comment 46 ABCA3 p.Glu690Lys 2068 G>A, (p.E690K) 17 23 24 TAT A GC GGG CTG ACA TCC 252627 Figure 1. ABCA3 exons 25-26 aberrant splicing. Login to comment 52 ABCA3 p.Glu690Lys 2068 A (p.E690K) eliminates Ban II site Size markers Ban II: + + Retained intron-specific amplicon Normal splice-specific amplicon Larger (645 bp) product indicates A at position 2068 586 bp 645 bp G A BanII sites 1872 bp 1865 bp P-intron P3-WT-span P1 P1 Figure 2. Transcript-specific RT-PCR and Ban II restriction endonuclease digest. Login to comment 53 ABCA3 p.Glu690Lys ABCA3 cDNA showing allele with inserted intronic sequence corresponding to the non-p.E690K allele. Login to comment 54 ABCA3 p.Glu690Lys Using a BanII restriction digest, the known p.E690K mutation eliminates a restriction site in the -normally spliced amplicon, generating a larger product (645 bp vs. 586 bp), -confirming the intronic insertion is in trans to the previously identified mutation. Login to comment 80 ABCA3 p.Glu690Lys ABCA3 p.Glu690Lys ABCA3 p.Leu941Pro ABCA3 p.Leu941Pro ABCA3 p.Leu212Met ABCA3 p.Leu212Met However, the mechanism to account for this larger aberrant transcript with an additional 249 bases Table 1. Characteristics of subjects with one ABCA3 mutation Patient Ethnicity Presentation Allele 1 mutation Allele 2 mutation Findings consistent with ABCA3 deficiency Outcome A Caucasian Newborn, RDS p.E690K IVS25-98T Case patient; lung histopathology and EM Died B Caucasian RDS p.L941P IVS25-98T Family history of sibling with fatal RDS Died C Caucasian 8 y/old, ILD L212M ? Login to comment 82 ABCA3 p.Glu1325Lys Family history of two siblings with fatal RDS, lung histopathology, and EM Died E,F Caucasian Newborn, RDS p.E1325K ? Login to comment 83 ABCA3 p.Arg43Cys ABCA3 p.Ala1070Thr Died G Hispanic 2 mo, ILD p.R43C IVS25-98T Lung histopathology and EM Lung transplant H Hispanic Newborn, RDS p.A1070T ? Login to comment 84 ABCA3 p.Arg280Cys ABCA3 p.Arg43His Mutation associated with disease in other patients, lung histopathology Alive with ILD I Caucasian Newborn, RDS p.R43H IVS25-98T Mutation associated with disease in other patients, lung histopathology Alive with ILD J African American ILD p.R280C ? Login to comment 85 ABCA3 p.Asn1418Ser Mutation associated with disease in other patients, lung histopathology Alive with ILD K Caucasian ILD p.N1418S ? Login to comment 116 ABCA3 p.Glu690Lys Genetic testing performed through the Johns Hopkins University DNA Diagnostic Lab revealed normal SFTPB and SFTPC gene sequences but a heterozygous mutation in ABCA3 (Nt2068G->A, p.E690K) that had previously been identified as disease causing (22,23). Login to comment 135 ABCA3 p.Glu690Lys Specific primers were created to generate amplicons expanding from the previously known mutation (p.E690K at exon 17) to the area of interest in intron 25. Login to comment