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. 1994 Dec;106(4):1325–1333. doi: 10.1104/pp.106.4.1325

The plasma membrane of Arabidopsis thaliana contains a mercury-insensitive aquaporin that is a homolog of the tonoplast water channel protein TIP.

M J Daniels 1, T E Mirkov 1, M J Chrispeels 1
PMCID: PMC159670  PMID: 7846153

Abstract

Plant cells contain proteins that are members of the major intrinsic protein (MIP) family, an ancient family of membrane channel proteins characterized by six membrane-spanning domains and two asparagine-proline-alanine (NPA) amino acid motifs in the two halves of the protein. We recently demonstrated that gamma-TIP, one of the MIP homologs found in the vacuolar membrane of plant cells, is an aquaporin or water channel protein (C. Maurel, J. Reizer, J.I. Schroeder, M.J. Chrispeels [1993] EMBO J 12: 2241-2247). RD28, another MIP homolog in Arabidopsis thaliana, was first identified as being encoded by a turgor-responsive transcript. To find out if RD28 is a water channel protein, rd28 cRNA was injected into Xenopus laevis oocytes. Expression of RD28 caused a 10- to 15-fold increase in the osmotic water permeability of the oocytes, indicating that the protein creates water channels in the plasma membrane of the oocytes and is an aquaporin just like its homology gamma-TIP. Although RD28 has several cysteine residues, its activity is not inhibited by mercury, and in this respect it differs from gamma-TIP and all but one of the mammalian water channels that have been described. Introduction of a cysteine residue next to the second conserved NPA motif creates a mercury-sensitive water channel, suggesting that this conserved loop is critical to the activity of the protein. Antibodies directed at the C terminus of RD28 were used in combination with a two-phase partitioning method to demonstrate that RD28 is located in the plasma membrane. The protein is present in leaves and roots of well-watered plants, suggesting that its presence in plants does not require a specific desiccation regime. These results demonstrate that plant cells contain constitutively expressed aquaporins in their plasma membranes (RD28), as well as in their tonoplasts (gamma-TIP).

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Selected References

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  1. Fray R. G., Wallace A., Grierson D., Lycett G. W. Nucleotide sequence and expression of a ripening and water stress-related cDNA from tomato with homology to the MIP class of membrane channel proteins. Plant Mol Biol. 1994 Feb;24(3):539–543. doi: 10.1007/BF00024122. [DOI] [PubMed] [Google Scholar]
  2. Fushimi K., Uchida S., Hara Y., Hirata Y., Marumo F., Sasaki S. Cloning and expression of apical membrane water channel of rat kidney collecting tubule. Nature. 1993 Feb 11;361(6412):549–552. doi: 10.1038/361549a0. [DOI] [PubMed] [Google Scholar]
  3. Guerrero F. D., Crossland L. Tissue-specific expression of a plant turgor-responsive gene with amino acid sequence homology to transport-facilitating proteins. Plant Mol Biol. 1993 Mar;21(5):929–935. doi: 10.1007/BF00027125. [DOI] [PubMed] [Google Scholar]
  4. Guerrero F. D., Jones J. T., Mullet J. E. Turgor-responsive gene transcription and RNA levels increase rapidly when pea shoots are wilted. Sequence and expression of three inducible genes. Plant Mol Biol. 1990 Jul;15(1):11–26. doi: 10.1007/BF00017720. [DOI] [PubMed] [Google Scholar]
  5. Höfte H., Hubbard L., Reizer J., Ludevid D., Herman E. M., Chrispeels M. J. Vegetative and Seed-Specific Forms of Tonoplast Intrinsic Protein in the Vacuolar Membrane of Arabidopsis thaliana. Plant Physiol. 1992 Jun;99(2):561–570. doi: 10.1104/pp.99.2.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Johnson K. D., Höfte H., Chrispeels M. J. An intrinsic tonoplast protein of protein storage vacuoles in seeds is structurally related to a bacterial solute transporter (GIpF). Plant Cell. 1990 Jun;2(6):525–532. doi: 10.1105/tpc.2.6.525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Kammerloher W., Fischer U., Piechottka G. P., Schäffner A. R. Water channels in the plant plasma membrane cloned by immunoselection from a mammalian expression system. Plant J. 1994 Aug;6(2):187–199. doi: 10.1046/j.1365-313x.1994.6020187.x. [DOI] [PubMed] [Google Scholar]
  8. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Maurel C., Reizer J., Schroeder J. I., Chrispeels M. J., Saier M. H., Jr Functional characterization of the Escherichia coli glycerol facilitator, GlpF, in Xenopus oocytes. J Biol Chem. 1994 Apr 22;269(16):11869–11872. [PubMed] [Google Scholar]
  10. Maurel C., Reizer J., Schroeder J. I., Chrispeels M. J. The vacuolar membrane protein gamma-TIP creates water specific channels in Xenopus oocytes. EMBO J. 1993 Jun;12(6):2241–2247. doi: 10.1002/j.1460-2075.1993.tb05877.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Preston G. M., Agre P. Isolation of the cDNA for erythrocyte integral membrane protein of 28 kilodaltons: member of an ancient channel family. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11110–11114. doi: 10.1073/pnas.88.24.11110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Preston G. M., Carroll T. P., Guggino W. B., Agre P. Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein. Science. 1992 Apr 17;256(5055):385–387. doi: 10.1126/science.256.5055.385. [DOI] [PubMed] [Google Scholar]
  13. Preston G. M., Jung J. S., Guggino W. B., Agre P. The mercury-sensitive residue at cysteine 189 in the CHIP28 water channel. J Biol Chem. 1993 Jan 5;268(1):17–20. [PubMed] [Google Scholar]
  14. Reizer J., Reizer A., Saier M. H., Jr The MIP family of integral membrane channel proteins: sequence comparisons, evolutionary relationships, reconstructed pathway of evolution, and proposed functional differentiation of the two repeated halves of the proteins. Crit Rev Biochem Mol Biol. 1993;28(3):235–257. doi: 10.3109/10409239309086796. [DOI] [PubMed] [Google Scholar]
  15. Sandal N. N., Marcker K. A. Soybean nodulin 26 is homologous to the major intrinsic protein of the bovine lens fiber membrane. Nucleic Acids Res. 1988 Oct 11;16(19):9347–9347. doi: 10.1093/nar/16.19.9347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Shagan T., Bar-Zvi D. Nucleotide sequence of an Arabidopsis thaliana turgor-responsive cDNA clone encoding TMP-A, a transmembrane protein containing the major intrinsic protein motif. Plant Physiol. 1993 Apr;101(4):1397–1398. doi: 10.1104/pp.101.4.1397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Smith D. W. A complete, yet flexible, system for DNA/protein sequence analysis using VAX/VMS computers. Comput Appl Biosci. 1988 Mar;4(1):212–212. doi: 10.1093/bioinformatics/4.1.212. [DOI] [PubMed] [Google Scholar]
  18. Vallee B. L., Ulmer D. D. Biochemical effects of mercury, cadmium, and lead. Annu Rev Biochem. 1972;41(10):91–128. doi: 10.1146/annurev.bi.41.070172.000515. [DOI] [PubMed] [Google Scholar]
  19. Verbavatz J. M., Brown D., Sabolić I., Valenti G., Ausiello D. A., Van Hoek A. N., Ma T., Verkman A. S. Tetrameric assembly of CHIP28 water channels in liposomes and cell membranes: a freeze-fracture study. J Cell Biol. 1993 Nov;123(3):605–618. doi: 10.1083/jcb.123.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Voelker T. A., Herman E. M., Chrispeels M. J. In vitro mutated phytohemagglutinin genes expressed in tobacco seeds: role of glycans in protein targeting and stability. Plant Cell. 1989 Jan;1(1):95–104. doi: 10.1105/tpc.1.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yamamoto Y. T., Taylor C. G., Acedo G. N., Cheng C. L., Conkling M. A. Characterization of cis-acting sequences regulating root-specific gene expression in tobacco. Plant Cell. 1991 Apr;3(4):371–382. doi: 10.1105/tpc.3.4.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Zeidel M. L., Ambudkar S. V., Smith B. L., Agre P. Reconstitution of functional water channels in liposomes containing purified red cell CHIP28 protein. Biochemistry. 1992 Aug 25;31(33):7436–7440. doi: 10.1021/bi00148a002. [DOI] [PubMed] [Google Scholar]
  23. Zhang R. B., Verkman A. S. Water and urea permeability properties of Xenopus oocytes: expression of mRNA from toad urinary bladder. Am J Physiol. 1991 Jan;260(1 Pt 1):C26–C34. doi: 10.1152/ajpcell.1991.260.1.C26. [DOI] [PubMed] [Google Scholar]

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