ABSTRACT
The PIP gene, localized in the 7q34 region that contains a number of fragile sites such as FRA 7H and FRA TI, codes for gp17/PIP, a protein secreted by breast apocrine tumors. We analyzed the integrity of this gene in 20 tumors of the urogenital tract. We found rearranged EcoRI fragments in 5 of 15 primary prostate carcinomas. No rearrangement was found in normal prostates derived from five patients undergoing prostatocystectomy during treatment of bladder cancers. By Southern blot hybridization with PIP gene exon-specific probes, the rearrangements were mapped at or near the 3' end of the gene. These abnormalities were found, not only in the neoplastic cells invading the prostatic tissues, but also in seminal vesicles without histologic tumoral features. These data suggest a critical role of the PIP gene or neighboring genes in prostate cancer.
Subject(s)
Apolipoproteins , Biomarkers, Tumor/genetics , Carrier Proteins/genetics , Glycoproteins , Membrane Transport Proteins , Polymorphism, Restriction Fragment Length , Prostatic Neoplasms/genetics , Apolipoproteins D , Blotting, Southern , Carcinoma/genetics , DNA, Neoplasm/chemistry , Deoxyribonuclease EcoRI/chemistry , Humans , Male , Restriction Mapping , Translocation, Genetic/genetics , Urinary Bladder Neoplasms/geneticsABSTRACT
We have previously isolated from human seminal plasma a CD4 ligand, the gp17 glycoprotein, which shares sequence identity with three previously identified proteins: secretory actin-binding protein (SABP) from seminal plasma, gross-cystic-disease fluid protein-15 (GCDFP-15) and prolactin-inducible protein (PIP) from breast tumor cells. Functions of these glycoproteins are unknown. To further characterize the physical interaction between gp17 and CD4 we used surface plasmon resonance and demonstrated that gp17-CD4 binding affinity is high. Competition experiments indicated that gp17 interferes with human immunodeficiency virus (HIV) envelope protein/CD4 binding, although it binds to a site distinct from but close to the gp120-binding site. We observed moreover that gp17 inhibits syncytium formation between transfected cells expressing the wild-type HIV-1 envelope glycoprotein and CD4, respectively. Our results suggest that gp17, which may function as an immunomodulatory CD4-binding factor playing a role at insemination, may also play a role in controlling HIV spread in the sexual tract.
Subject(s)
Apolipoproteins , CD4 Antigens/metabolism , Carrier Proteins/chemistry , Giant Cells/drug effects , Glycoproteins/chemistry , HIV Envelope Protein gp120/metabolism , HIV-1/drug effects , Membrane Transport Proteins , Semen/chemistry , Apolipoproteins D , Binding, Competitive , Carrier Proteins/pharmacology , Glycoproteins/pharmacology , HeLa Cells , Humans , Kinetics , Recombinant Proteins/metabolismSubject(s)
Apolipoproteins , CD4 Antigens/metabolism , Carrier Proteins/genetics , Glycoproteins/genetics , Membrane Transport Proteins , Neoplasm Proteins/genetics , Seminal Vesicles/chemistry , Amino Acid Sequence , Apolipoproteins D , Base Sequence , Blotting, Northern , Carrier Proteins/metabolism , Escherichia coli , Gene Library , Glycoproteins/metabolism , Humans , Male , Molecular Sequence Data , Neoplasm Proteins/metabolism , RNA, Messenger/analysisABSTRACT
Although the HIV gp120 binding site of CD4 is well characterized, its interaction site with HLA class II molecules is still controversial. Sixty-seven mutations within the four extracellular domains of CD4 were examined for their effects on cell surface expression and conformational changes and for adhesion of HLA class II-expressing B lymphocytes and HIV gp120 binding to CD4-transfected COS cells. Mutations within the gp120 binding site affected both assays similarly, indicating that the two sites fully overlap. A few additional substitutions of residues mapping on the same face of domains 1 and 2 induced decreased rosette formation. Molecular modeling studies indicated that this is likely to be the consequence of conformational changes induced by the mutations. Thus, CD4 appears to interact with HLA class II molecules mainly through the HIV gp120 binding site and possibly through a second minor interaction site mapping on the same face of the molecule.