A 25-kb deletion in the 5′ region of the cytochrome b558 heavy chain gene (CYBB) in a patient with X-linked chronic granulomatous disease

Abstract

We performed molecular genetic analyses of the family of a boy suffering from chronic granulomatous disease (CGD) after immunocytochemically confirming him and his mother to be an X-linked CGD patient and a mosaic carrier, respectively. Southern blot hybridization using cDNA for the cytochrome b 558 heavy chain gene (CYBB) as a probe showed that the patient had a deletion in the 5′ region of the CYBB and his phenotypically normal mother was heterozygous for this deletion. Polymerase chain reaction analyses of all 13 exons of the patient's CYBB gene demonstrated that the deletion extends from exon 7 or neighboring introns to 5′ upstream. The length of the deletion was determined by pulsed-field gel electrophoresis and Southern blotting of genomic DNA using CYBB cDNA and the genetic marker pERT55-5, centromeric to CYBB, as probes. Both probes recognized common SfiI-NotI fragments of 120 kb and 95 kb in normal individuals and the patient, respectively. These results revealed that the patient has a 25-kb deletion spanning from the middle of CYBB to 5′ upstream. This is the only report of a large 5′ deletion in CYBB and also the first observation that CYBB and pERT55-5 are within 120 kb in Xp21.

Nonhomologous Recombination between the Cytochrome b558Heavy Chain Gene (CYBB) and LINE-1 Causes an X-Linked Chronic Granulomatous Disease

“…Altered bands (arrow) of StuI-digested DNAs in the patient and his mother (P and M, respectively in Fig. 4A) were consistent with the findings reported previously (Faizunnessa et al, 1997). The patient's grandmother (G) and aunt (A) were genetically normal, as were his brother (B) and an unrelated normal person (C) (Fig.…”

“…Our previous study showed that this CGD patient has the 25-kb deletion upstream of exon 7 or neighboring introns of CYBB (Faizunnessa et al, 1997). To determine breakpoints in his CYBB, the patient's genomic DNA library containing 10-to 15-kb Sau3AI fragments was screened with the labeled 5Ј half of the CYBB cDNA as a probe.…”

“…In a previous study, we determined the size (25 kb) of a deletion in an X-linked CGD patient by pulsed-field gel electrophoresis (Faizunnessa et al, 1997). In this study we determined nucleotide sequences around the deletion junction in this CGD patient and identified its 5Ј and 3Ј breakpoints.…”

Statistical and mutational analysis of chronic granulomatous disease in Japan with special reference to gp91-phox and p22-phox deficiency

“…We have carried out a statistical analysis of 229 CGD patients from 195 families in Japan and the mutation analysis of an additional 28 independent gp91-phox deficient and five independent p22-phox deficient patients to those published previously from Japan (Ariga et al 1998;Azuma et al 1995;Faizunnessa et al 1997;Kaneda et al 1999;Okubo et al 1992;Suzuki et al 1998;Tsuda et al 1998;Yoshida et al 1998). Altogether, mutation analysis of a total of 48 out of 109 cases identified as gp91-phox deficiency (44%) and a total of five out of 16 cases of p22-phox deficiency in Japan (31%) has been accomplished.…”

“…The mutation analysis of the other 20 Japanese cases has been reported (Ariga et al 1998;Azuma et al 1995;Faizunnessa et al 1997;Kaneda et al 1999;Okubo et al 1992;Suzuki et al 1998;Tsuda et al 1998;Yoshida et al 1998).…”