Genomic Structure of the Human p47-phox (NCF1) Gene

Abstract

ABSTRACT:The cytosolic factor p47-phox, encoded by the NCF1 gene, is an essential component of the phagocyte NADPH-oxidase system. Upon activation of this multicomponent system, p47-phox translocates to the membrane and participates in the electron transfer from NADPH to molecular oxygen. A deficiency or absence of p47-phox is the most common autosomal form of chronic granulomatous disease (CGD). We have cloned and characterized the NCF1 gene from four bacteriophage clones, a P1 clone and genomic DNA from normal individuals. The gene is 15,236 base pairs long and includes 11 exons. It is 98.6% homologous in sequence to at least one pseudogene that maps to the same region of chromosome 7q11.23. Slightly more than half (50.37%) of the wild-type NCF1 gene consists of repetitive elements. In particular, the density of Alu sequences is high (1.4 Alu/kb); there are 21 Alu repeats interspersed through 10 introns. These findings are consistent with the observation that recombination events between the wild-type gene and its highly homologous pseudogenes account for the majority of potentially lethal mutations in p47-phox-deficient chronic granulomatous disease. Analysis of 1.96 kb of sequence 5Ј of the start of translation revealed a high homology (99.6%) between wild-type and pseudogene clones. Characterization of NCF1 establishes a foundation for detailed molecular analysis of p47-phox-deficient CGD patients as well as for the study of the regulation of the NCF1 gene and pseudogenes, both of which are present as full-length transcripts in normal individuals.

A founder effect for p47phox Trp193Ter chronic granulomatous disease in Kavkazi Jews

“…2) was performed by Sanger sequencing according to [5]; reference for the sequence was retrieved from [20]. Primers were developed on NCF1-specific points in the introns of NCF1 to amplify only NCF1-and not pseudo-genespecific sequences (Suppl.…”

A false-carrier state for the c.579G>A mutation in the NCF1 gene in Ashkenazi Jews

“…Sequencing of exon 4 and exon 7 of NCF1 was performed by Sanger sequencing according to Roos et al ,6 and reference for the sequence was retrieved from Chanock et al 7. Segregating SNPs between NCF1 and its pseudogenes showed that the PCR was NCF1 gene specific.…”

Contribution of genetic studies in rodent models of autoimmune arthritis to understanding and treatment of rheumatoid arthritis

“…Recent evidence implicates interferon-b as a mediator of NAPDH-enhancing disease amelioration. 60 The human NCF1 gene has two pseudogene duplications, 61,62 and has proven difficult to study in humans.…”