Analysis of the promoter of mutated human whey acidic protein (WAP) gene.
Although whey acidic protein (WAP) has been identified in the milk of a range of species, it has been predicted that WAP is not secreted into human milk as a result of critical point mutations within the coding region. In the present study, we first investigated computationally the promoter region of mutated human WAP genes by comparing with those of other known WAP genes. Computational database analyses showed that the human WAP promoter region was highly conserved, as in other species with milk WAP. Next, we evaluated the activity of the human WAP promoter (2.6 kb) using a reporter gene assay. MCF-7 cells were stably transfected with the hWAP/hGH (human growth hormone) fusion gene, cultured on Matrigel, and treated with lactogenic hormones. Radioimmunoassay detected hGH in the culture medium, indicating that the human WAP promoter was responsible for the lactogenic hormones. The human WAP promoter was significantly more active in MCF-7 cells than the mouse WAP promoter (2.4 kb). The present results provide us with important information on the molecular evolution of milk protein genes.
Nukumi
Naoko
N
Laboratory of Applied Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Japan.
Seki
Mami
M
Iwamori
Tokuko
T
Yada
Tetsushi
T
Naito
Kunihiko
K
Tojo
Hideaki
H
D045744:Cell Line, Tumor; D003094:Collagen / Q000378:metabolism / Q000494:pharmacology; D019295:Computational Biology; D017124:Conserved Sequence; D030541:Databases, Genetic; D004338:Drug Combinations; D019143:Evolution, Molecular; D006801:Humans; D007797:Laminin / Q000378:metabolism / Q000494:pharmacology; D008894:Milk Proteins / Q000235:genetics; D008895:Milk, Human; D008957:Models, Genetic; D015233:Models, Statistical; D017354:Point Mutation; D011401:Promoter Regions, Genetic; D011509:Proteoglycans / Q000378:metabolism / Q000494:pharmacology; D011863:Radioimmunoassay; D017422:Sequence Analysis, DNA; D014162:Transfection
D004338:Drug Combinations; D007797:Laminin; D008894:Milk Proteins; D011509:Proteoglycans; C024471:whey acidic proteins; C059485:matrigel; D003094:Collagen