Molecular Cloning of Vps26a, Vps26b, Vps29, and Vps35 and Expression Analysis of Retromer Complex in Micro Pig

  • Kim, Ek-Yune (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Kim, Young-Hyun (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Ryu, Chung-Hun (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Lee, Jae-Woong (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Kim, Sang-Hyun (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Lee, Sang-Rae (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Kim, Myeong-Su (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Kim, Wan-Jun (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB)) ;
  • Lim, Jeong-Mook (Department of Agricultural Biotechnology, Division of Animal Genetic Engineering, Seoul National University) ;
  • Chang, Kyu-Tae (National Primate Research Center (NPRC), Korea Research Institute of Biotechnology (KRIBB))
  • Published : 2008.03.31

Abstract

Members of the Vps (Vacuolar protein sorting) protein family involved in the formation of the retromer complex have been discovered in a variety of species such as yeast, mouse, and human. A mammalian retromer complex is composed of Vps26, Vps29, and Vps35 proteins and plays and important role in cation-independent mannose-6-phosphate receptor retrieval from the endosome to the trans-Golgi network. In this study, we have identified the full-length sequences of the retromer components of Vps26, Vps29, and Vps35 in micro pigs. The cDNA sequences of these retromer components have been determined and the result showed there is 99% homology among the component counterparts from mouse, micro pigs, and humans. In addition, the retromer complexes formed with hetero-components were found in the brain of micro pigs. Based on above results, we suggest mammalian Vps components are well conserved in micro pigs.

Keywords

References

  1. Bachhawat AK, Suhan J, Jones EW (1994): The yeast homolog of H beta 58, a mouse gene essential for embryogenesis, performs a role in the delivery of proteins to the vacuole. Genes Dev 8:1379-1387 https://doi.org/10.1101/gad.8.12.1379
  2. Bonifacino JS, Lippincott-Schwartz J (2003): Coat proteins: shaping membrane transport. Nat Rev Mol Cell Biol 4:409-414 https://doi.org/10.1038/nrm1099
  3. Bradford MM (1976): A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254 https://doi.org/10.1016/0003-2697(76)90527-3
  4. Carlton J, Bujny M, Peter BJ, Oorschot VM, Rutherford A, Mellor H, Klumperman J, McMahon HT, Cullen PJ (2004): Sorting nexin-1 mediates tubular endosome-to-TGN transport through coincidence sensing of high-curvature membranes and 3-phosphoinositides. Curr Biol 14:1791-1800 https://doi.org/10.1016/j.cub.2004.09.077
  5. Collins BM, Skinner CF, Watson PJ, Seaman MN, Owen DJ (2005): Vps29 has a phosphoesterase fold that acts as a protein interaction scaffold for retromer assembly. Nat Struct Mol Biol 12:594-602 https://doi.org/10.1038/nsmb954
  6. Collins BM, Norwood SJ, Kerr MC, Mahony D, Seaman MN, Teasdale RD, Owen DJ (2008): Structure of Vps26B and mapping of its interaction with the retromer protein complex. Traffic 9:366-379 https://doi.org/10.1111/j.1600-0854.2007.00688.x
  7. Emr SD, Vassarotti A, Garrett J, Geller BL, Takeda M, Douglas MG (1986): The amino terminus of the yeast F1-ATPase beta-subunit precursor functions as a mitochondrial import signal. J Cell Biol 102:523-533 https://doi.org/10.1083/jcb.102.2.523
  8. Gietz, D, Jean A, Woods RA, Schiestl RH (1992): Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425 https://doi.org/10.1093/nar/20.6.1425
  9. Gokool S, Tattersall D, Reddy JV, Seaman MN (2007): Identification of a conserved motif required for Vps-35p/Vps26p interaction and assembly of the retromer complex. Biochem J 408:287-295 https://doi.org/10.1042/BJ20070555
  10. Haft CR, de la Luz Sierra M, Bafford R, Lesniak MA, Barr VA, Taylor SI (2000): Human orthologs of yeast vacuolar protein sorting proteins Vps26, 29, and 35:assembly into multimeric complexes. Mol Biol Cell 11:4105-4116 https://doi.org/10.1091/mbc.11.12.4105
  11. Kerr MC, Bennetts JS, Simpson F, Thomas EC, Flegg C, Gleeson PA, Wicking C, Teasdale RD (2005): A novel mammalian retromer component, Vps26B. Traffic 6:991-1001 https://doi.org/10.1111/j.1600-0854.2005.00328.x
  12. Kim E, Yamashita E, Nakanishi T, Park K, Kimura M, Kashiwabara S, Baba T (2006): Mouse sperm lacking ADAM1b/ADAM2 fertilin can fuse with the egg plasma membrane and effect fertilization. J Biol Chem 281:5634-5639 https://doi.org/10.1074/jbc.M510558200
  13. Kurten RC, Cadena DL, Gill GN (1996): Enhanced degradation of EGF receptors by a sorting nexin, SNX1. Science 272:1008-1010 https://doi.org/10.1126/science.272.5264.1008
  14. Lee JJ, Radice G, Perkins CP, Costantini F (1992): Identificatio and characterization of a novel, evolutionarily conserved gene disrupted by the murine H beta 58 embryonic lethal transgene insertion. Development 115:227-288
  15. Mizuno E, Nakamura M, Agemura A, Kusumoto A, Ichiba M, Kurano Y, Muroya S, Sano A (2007): Brain-specific transcript variants of 5' and 3' ends of mouse VPS13A and VPS13C
  16. Nothwehr SF, Ha SA, Bruinsma P (2003): Sorting of Yeast membrane proteins into an dndosome-to Golgi pathwy involves direct interaction of their cytosolic domains with Vps35p. J Cell Biol 4:409-414
  17. Prasad BC, Clark SG (2006): Wnt signaling establishes anteroposterior neuronal polarity and requires retromer in C. elegans. Development 133:1757-1766 https://doi.org/10.1242/dev.02357
  18. Reddy JV, Seaman MN (2001): Vps26p, a component of retromer, directs the interactions of Vps35p in endosome-to-Golgi retrieval. Mol Biol Cell 12:3242-3256 https://doi.org/10.1091/mbc.12.10.3242
  19. Schwarz DG, Griffin CT, Schneider EA, Yee D, Magnuson T (2002): Genetic analysis of sorting nexins 1 and 2 reveals a redundant and essential function in mice. Mol Biol Cell 13:3588-3600 https://doi.org/10.1091/mbc.E02-03-0145
  20. Seaman MN. McCaffery JM, Emr SD (1998): A membrane coat complex essential for endosome-to-Golgi retrograde transport in yeast. J Cell Biol 142:665-681 https://doi.org/10.1083/jcb.142.3.665
  21. Seaman MN (2004): Cargo-selective endosomal sorting for retrieval to the Golgi requires retromer. J Cell Biol 165:111-122 https://doi.org/10.1083/jcb.200312034
  22. Verges M, Sebastian I, Mostov KE (2007): Phosphoinositide 3-kinase regulates the role of retromer in transcytosis of the polymeric immunoglobulin receptor. Exp Cell Res 313:707-718 https://doi.org/10.1016/j.yexcr.2006.11.010