The tissue distribution of the mRNAs for a number of salivary proteins [proline-rich proteins (PRPs), statherin, cystatins, and the histatins] has been examined in humans and macaques in order to investigate their possible functions and tissue-specific regulation. We have shown that PRP RNAs (0.8-1.5 kb) are expressed in human and rhesus parotid and submandibular glands, and in the human bronchus. The genes for the acidic and basic PRPs are differentially regulated in these tissues. RNAs for acidic PRPs are predominantly expressed in the submandibular gland, for basic PRPs in the respiratory tract, and for both acidic and basic PRPs in the parotid gland. Protein studies of secretions from these tissues confirm the RNA results. Statherin RNA (0.65 kb) was detected in human and rhesus parotid and submandibular glands and the human bronchus, as well as in rhesus lacrimal glands. Statherin was found by tissue immunoperoxidase staining in the serous cells of respiratory tract submucosal glands, which is the same location for the synthesis of PRPs. Several cystatin RNAs (0.8-1.3 kb) were differentially expressed in human parotid glands, submandibular glands, and the bronchus, and in lacrimal glands from both rhesus and cynomolgus macaques. RNAs (0.6 kb) for the histatins were found only in parotid and submandibular glands. Thus, it appears that PRPs, statherin, and cystatins may play a broader role in the physiology of biological fluids and secretions than previously suspected, since they are found in secretions other than saliva. However, the functions of the histatins are restricted to saliva. These studies also pose some interesting questions regarding the differential expression of these genes in a variety of secretory tissues.
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References
1.
Abrahamson, M.; Barrett, A.J.; Salvesen, G.; and Grubb, A. (1986): Isolation of Six Cysteine Proteinase Inhibitors from Human Urine. Their Physicochemical and Enzyme Kinetic Properties and Concentrations in Biological Fluids, J Biol Chem261:11282-11289.
2.
Abrahamson, M.; Grubb, A.; Olafsson, I.; and Lund-Wall, Å. ( 1987): Molecular Cloning and Sequence Analysis of cDNA Coding for the Precursor of the Human Cysteine Proteinase Inhibitor Cystatin C, FEBS Lett216:229-233.
3.
Al-Hashimi, I.; Dickinson, D.P.; and Levine, M.J. (1988): Purification, Molecular Cloning, and Sequencing of Salivary Cystatin SA-I, J Biol Chem263:9381-9387.
4.
Azen, E.A. (1972): Genetic Polymorphism of Basic Proteins from Parotid Saliva, Science176:673-674.
Azen, E.A. (1978): Phosphorylation of Proline-rich, Double Band, Acidic and Post-Pb Proteins of Human Saliva, Arch Oral Biol23:1173-1176.
7.
Azen, E.A. (1989): Genetic Protein Polymorphisms of Human Saliva . In: Clinical Chemistry of Human Saliva, Vol. 1, J. Tenovuo, Ed., Boca Raton, FL: CRC Press, Inc., Chap. 7, pp. 161-195.
8.
Azen, E.A. and Denniston, C. (1981): Genetic Polymorphism of PIF (Parotid Isoelectric Focusing Variant) Proteins with Linkage to the PPP (Parotid Proline-Rich Protein) Gene Complex, Biochem Genet19:475-485.
9.
Azen, E.A.; Goodman, P.A.; and Lalley, P.A. (1985): Human Salivary Proline-Rich Protein Genes on Chromosome 12, Am J Hum Genet37:418-424.
10.
Azen, E.A.; Leutenegger, W.; and Peters, E.H. (1978): Evolutionary and Dietary Aspects of Salivary Basic (Pb) and Post Pb (PPb) Proteins in Anthropoid Primates, Nature273:775-778.
11.
Azen, E.A.; Lyons, K.M.; McGonigal, T.; Barrett, N.L.; Clements, L.S.; Maeda, N.; Vanin, E.F.; Carlson, D.; and Smithies, 0. ( 1984): Clones from the Human Gene Complex Coding for Salivary Proline-Rich Proteins, Proc Natl Acad Sci USA81:5561-5565.
12.
Azen, E.A. and Maeda, N. (1988): Molecular Genetics of Human Salivary Proteins and their Polymorphisms. In: Advances in Human Genetics, Vol. 17, H. Harris and K. Hirschhorn, Eds., New York, NY: Plenum Press, pp. 141-199.
13.
Azen, E.A. and Yu, P.-L. (1984): Genetic Polymorphism of CON 1 and CON 2 Salivary Proteins Detected by Immunologic and Concanavalin A Reactions on Nitrocellulose with Linkage of CON1 and CON2 Genes to the SPC (Salivary Protein Gene Complex) , Biochem Genet22:1-19.
14.
Baum, B.J.; Bird, J.L.; and Longton, R.W. (1977): Polyacrylamide Gel Electrophoresis of Human Salivary Histidine-rich Polypeptides, J Dent Res56:1115-1118.
15.
Bennick, A. (1982): Salivary Proline-rich Proteins, Mol Cell Biochem45:83-99.
16.
Bennick, A.; Chau, G.; Goodlin, R.; Abrams, S.; Tus-Tian, D.; and Madapallimattam, G. (1983): The Role of Human Salivary Acidic Proline-rich Proteins in the Formation of Acquired Dental Pellicle in vivo and their Fate after Adsorption to the Human Enamel Surface, Arch Oral Biol28:19-27.
17.
Chirgwin, J.M.; Przybyla, A.E.; Macdonald, R.J.; and Rutter, W.J. (1979): Isolation of Biologically Active Ribonucleic Acid from Sources Enriched in Ribonuclease, Biochemistry18:5294-5299.
18.
Church, G.M. and Gilbert, W. (1984): Genomic Sequencing, Proc Natl Acad Sci USA81:1991-1995.
19.
Curby, W.A. (1953): Device for Collection of Human Parotid Saliva , J Lab Clin Med41:493-496.
20.
Dickinson, D.P.; Ridall, A.L.; and Levine, M.J. (1987): Human Submandibular Gland Statherin and Basic Histidine-rich Peptide are Encoded by Highly Abundant mRNA's Derived from a Common Ancestral Sequence, Biochem Biophys Res Commun149:784-790.
21.
Feinberg, A.P. and Vogelstein, B. (1983): A Technique for Radiolabeling DNA Restriction Endonuclease Fragments to High Specific Activity, Anal Biochem132:6-13.
22.
Ghiso, J.; Jensson, O.; and Frangione, B. (1986): Amyloid Fibrils in Hereditary Cerebral Hemorrhage with Amyloidosis of Icelandic Type is a Variant of γ-trace Basic Protein (Cystatin C), Proc Natl Acad Sci USA83:2974-2978.
23.
Gibbons, R.J. and Hay, D.I. (1988): Human Salivary Acidic Proline-Rich Proteins and Statherin Promote the Attachment of Actinomyces viscosus LY7 to Apatitic Surfaces , Infect Immun56 :439-445.
24.
Green, G.D.J.; Kembhavi, A.A.; Davies, M.E.; and Barrett, A.J. (1984): Cystatin-Like Cysteine Proteinase Inhibitors from Human Liver, Biochem J218:939-946.
25.
Hagerman, A.E. and Butler, L.G. (1981): The Specificity of
26.
Proanthocyanidin-Protein Interactions, J Biol Chem256:4494-4497. Hatton, M.N.; Loomis, R.E.; Levine, M.J.; and Tabak, L.A. (1985): Masticatory Lubrication. The Role of Carbohydrate in the Lubricating Property of a Salivary Glycoprotein-Albumin Complex, Biochem J230:817-820.
27.
Hay, D.I.; Smith, D.J.; Schluckebier, S.K.; and Moreno, E.C. (1984): Relationship Between Concentration of Human Salivary Statherin and Inhibition of Calcium Phosphate Precipitation in Stimulated Human Parotid Saliva, J Dent Res63:857-863.
28.
Isemura, S.; Saitoh, E.; Ito, S.; Isemura, M.; and Sanada, K. (1984): Cystatin S: a Cysteine Proteinase Inhibitor of Human Saliva, J Biochem96:1311-1314.
29.
Isemura, S.; Saitoh, E.; Sanada, K.; Isemura, M.; and Ito, S. (1986): Cystatin S and the Related Cysteine Proteinase Inhibitors in Human Saliva. In: Cysteine Proteinases and Their Inhibitors, V. Turk, Ed., Berlin: de Gruyter, pp. 497-505.
30.
Ito, S.; Isemura, S.; Saitoh, E.; Sanada, K.; Suzuki, T.; and Shibata, A. (1983): Immunohistochemical Demonstration of Salivary Proline-Rich Peptide P-C Like Immunoreactivity in Human Pancreatic B-Cells , Acta Endocrinol103:544-551.
31.
Ito, S.; Suzuki, T.; Momotsu, T.; Isemura, S.; Saitoh, E.; Sanada, K.; and Shibata, A. (1985): Presence of Salivary Protein C and Salivary Peptide P-C-like Immunoreactivity in the Laryngo-Tracheo-Bronchial Glands, Acta Endocrinol108 :130-134.
32.
Kim, H.-S. and Maeda, N. (1986): Structures of Two HaeIII-type Genes in the Human Salivary Proline-rich Protein Multigene Family, J Biol Chem261:6712-6718.
33.
Kousvelari, E.E.; Baratz, R.S.; Burke, B.; and Oppenheim, F.G. (1980): Immunochemical Identification and Determination of Proline-rich Proteins in Salivary Secretions, Enamel Pellicle, and Glandular Tissue Specimens, J Dent Res59 :1430-1438.
34.
Lyons, K.M.; Azen, E.A.; Goodman, P.A.; and Smithies, O. (1988): Many Protein Products from a Few Loci: Assignment of Human Salivary Proline-Rich Proteins to Specific Loci, Genetics120:255-265.
35.
Mackay, B.J.; Denepitiya, L.; Iacono, V.J.; Krost, S.B.; and Pollock, J.J. (1984): Growth-inhibitory and Bactericidal Effects of Human Parotid Salivary Histidine-Rich Polypeptides on Streptococcus mutans , Infect Immun44:695-701.
36.
Maeda, N. (1985): Inheritance of the Human Salivary Proline-rich Proteins: a Reinterpretation in Terms of Six Loci Forming Two Subfamilies , Biochem Genet23:455-464.
37.
Maeda, N.; Kim, H.-S.; Azen, E.A.; and Smithies, O. (1985): Differential RNA Splicing and Post-Translational Cleavages in the Human Salivary Proline-Rich Protein Gene System, J Biol Chem260:11123-11130.
38.
Mamula, P.W.; Heerema, N.A.; Palmer, C.G.; Lyons, K.M.; and Karn, R.C. (1985): Localization of the Human Salivary Proteins Complex (SPC) to Chromosome Band - 12p13.2, Cytogenet Cell Genet39:279-284.
39.
McMaster, G.K. and Carmichael, G.G. (1977): Analysis of Single- and Double-Stranded Nucleic Acids on Polyacrylamide and Agarose Gels by Using Glyoxal and Acridine Orange, Proc Natl Acad Sci USA74:4835-4838.
40.
Menhansho, H.; Butler, L.G.; and Carlson, D.M. (1987): Dietary Tannins and Salivary Proline-Rich Proteins: Interactions, Induction and Defense Mechanisms. In: Annual Review of Nutrition, Vol. 7, R.E. Olson , E. Beutler, and H.P. Broquist, Eds., Palo Alto, CA: Annual Reviews, pp. 423-440.
41.
Minakata, K. and Asano, M. (1984): New Protein Inhibitors of Cysteine Proteinases in Human Saliva and Salivary Glands, Hoppe-Seyler's Z Physiol Chem365:399-403.
42.
Muller-Esterl, W.; Fritz, H.; Kellermann, J.; Lott-Speich, F.; Machleidt, W.; and Turk, V. (1986): The Mammalian Cysteine Proteinase Inhibitors. Structural Diversity and Evolutionary Origin. In: Cysteine Proteinases and Their Inhibitors , V. Turk, Ed., Berlin: de Gruyter, pp. 369-392.
43.
Oppenheim, F.G.; Xu, T.; McMillian, F.M.; Levitz, S.M.; Diamond, R.D.; Offner, G.D.; and Troxler, R.F. (1988): Histatins, a Novel Family of Histidine-rich Proteins in Human Parotid Secretion. Isolation, Characterization, Primary Structure, and Fungistatic Effects on Candida albicans, J Biol Chem263 :7472-7477.
44.
Oppenheim, F.G.; Yang, Y.-C.; Diamond, R.D.; Hyslop, D.; Offner, G.D.; and Troxler, R.F. (1986): The Primary Structure and Functional Characterization of the Neutral Histidine-rich Polypeptide from Human Parotid Secretion, J Biol Chem261:1177-1182.
45.
Pals, G. and Pronk, J.C. (1979): Genetic Variation in Parotid Basic Proteins (Pb) in the Bozo (Mali, West Africa), Hum Genet49:355-359.
46.
Peters, E.H. and Azen, E.A. (1977): Isolation and Partial Characterization of Human Parotid Basic Proteins, Biochem Genet15:925-946.
47.
Peters, E.H.; Goodfriend, T.; and Azen, E.A. (1977): Human Pb, Human Post-Pb, and Nonhuman Primate Pb Proteins: Immunological and Biochemical Relationships, Biochem Genet15:947-962.
48.
Pollock, J.J.; Denepitiya, L.; Mackay, B.J.; and Iacono, V.J. (1984): Fungistatic and Fungicidal Activity of Human Parotid Salivary Histidine-rich Polypeptides on Candida albicans, Infect Immun44:702-707.
49.
Sabatini, L.M. and Azen, E.A. (1989): Histatins, a Family of Salivary Histidine-rich Proteins, are Encoded by at Least Two Loci (Hisl and His2), Biochem Biophys Res Commun (in press).
50.
Sabatini, L.M.; Carlock, L.R.; Johnson, G.W.; and Azen, E.A. (1987): cDNA Cloning and Chromosomal Localization (4qll-13) of a Gene for Statherin, a Regulator of Calcium in Saliva, Am J Hum Genet41:1048-1060.
51.
Saitoh, E.; Isemura, S.; Sanada, K.; Kim, H.-S.; Smithies, O.; and Maeda, N. (1988): Cystatin Superfamily: Evidence that Family II Cystatin Genes are Evolutionarily Related to Family III Cystatin Genes, Biol Chem Hoppe-Seyler (Suppl)369:191-197.
52.
Saitoh, E.; Kim, H.-S.; Smithies, O.; and Maeda, N. (1987): Human Cysteine-Proteinase Inhibitors: Nucleotide Sequence Analysis of Three Members of the Cystatin Gene Family, Gene61:329-338.
53.
Shibata, S.; Nagata, K.; Nakamura, R.; Tsunemitsu, A.; and Misaki, A. (1980): Effect of Some Factors on Binding of Parotid Saliva Basic Glycoprotein to Oral Streptococci, J Periodontol51:499-504.
54.
Shomers, J.P.; Tabak, L.A.; Levine, M.J.; Mandel, I.D.; and Hay, D.I. (1982): Properties of Cysteine-containing Phosphoproteins from Human Submandibular-Sublingual Saliva, J Dent Res61:397-399.
55.
Vieira, J. and Messing, J. (1987): Production of Single-stranded Plasmid DNA, Methods Enzymol153:3-11.
56.
Warner, T.F. and Azen, E.A. (1984): Proline-rich Proteins are Present in Serous Cells of Submucosal Glands in the Respiratory Tract, Am Rev Respir Dis130:115-118.
