title: Cloning and sequencing of the 52K cathepsin D complementary deoxyribonucleic acid of MCF7 breast cancer cells and mapping on chromosome 11.
abstract: Two lambda gt11 libraries containing complementary DNAs from human breast cancer MCF7 cells were screened by expression with monoclonal antibodies to the secreted 52K protein and with a 36-mer oligonucleotide derived from the N-terminal amino acid sequence of the secreted 52K protein. Four overlapping clones were sequenced, and found to be extensively homologous to the cathepsin D of normal human kidney, except for 5-point mutations resulting in one amino acid change (Ala to Val) in the profragment of cathepsin D. Northern blot analysis showed the 2.2 kilobase (kb) cathepsin D mRNA to be induced by estradiol in MCF7 cells and produced constitutively at high levels in the estrogen-receptor-negative BT20 cell line. A simple restriction pattern consistent with the restriction map of cathepsin D cDNA was obtained in Southern blot analysis of MCF7 cell DNA. In situ hybridization of the 52K-9 cDNA probe on normal lymphocytes assigned the 52K cathepsin D gene at the extremity of the short arm of chromosome 11, in the p15 band, close to the H-ras gene and in the region whose deletion increases the risk of invasive breast cancer. We conclude that the estrogen induced 52K protein has the same sequence as normal pro-cathepsin D and we propose that the 52K protein correspond to the only pro-cathepsin D expressed in MCF7 cells.
authors.affiliation: Unité d'Endocrinologie Cellulaire et Moléculaire, (INSERM U 148), Montpellier, France.
authors.forename: P
authors.initials: P
authors.lastname: Augereau
authors.forename: M
authors.initials: M
authors.lastname: Garcia
authors.forename: M G
authors.initials: MG
authors.lastname: Mattei
authors.forename: V
authors.initials: V
authors.lastname: Cavailles
authors.forename: F
authors.initials: F
authors.lastname: Depadova
authors.forename: D
authors.initials: D
authors.lastname: Derocq
authors.forename: F
authors.initials: F
authors.lastname: Capony
authors.forename: P
authors.initials: P
authors.lastname: Ferrara
authors.forename: H
authors.initials: H
authors.lastname: Rochefort
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D002402:Cathepsin D / Q000235:genetics*; D002874:Chromosome Mapping; D002880:Chromosomes, Human, Pair 11; D003001:Cloning, Molecular; D006801:Humans; D014407:Tumor Cells, Cultured
chemical_list: D002402:Cathepsin D