Affiliation:
1. Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada L8N 3Z5
Abstract
The cDNA and genomic DNA for the µ3A subunit of the AP-3 (adaptor protein-3) complex were cloned from Chinese hamster cells. The AP-3 µ3A genes in Chinese hamster, human and mouse each comprise nine exons and eight introns, with all introns located in identical positions in the species studied. The AP-3 µ3A genes in these species are linked in a head-to-head fashion with the gene for the purine salvage pathway enzyme AK (adenosine kinase). These genes share the first exon, and a 512 bp fragment covering the intervening untranslated sequence has the characteristic of a CpG island promoter, and it effectively carried out transcription in both directions. Deletion studies indicate that this region contains both positive and negative regulatory elements affecting transcription of these genes. In comparison with the AP-3 µ3A gene (27 kb), the AK gene in human is very large (558 kb), with average exon and intron lengths of approx. 100 bp and 55.7 kb respectively. The ratio of non-coding to coding sequence in the human AK gene is >550, which is the highest reported for any gene. We also present evidence that a number of AK− mutants of Chinese hamster ovary cells contain large deletions that affect both of these genes. In addition to lacking part of the AK gene, two of these mutants also lacked all of the exons and introns corresponding to the AP-3 µ3A gene. These mutants should prove useful in elucidating the role of AP-3 µ3A in vesicle-mediated protein sorting – a process that is altered in Hermansky–Pudlak syndrome. Detailed phylogenetic analysis of the µ family of proteins presented here also provides insight into how different AP complexes are related and may have evolved.
Subject
Cell Biology,Molecular Biology,Biochemistry
Cited by
13 articles.
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