Abstract
The mechanism by which the mitochondrial large rRNA is involved in the restoration of the pole cell-forming ability in Drosophila embryos is still unknown. We identified a 15-ribonucleotide sequence which is conserved from the protobacterium Wolbachia to the higher eukaryotes in domain V of the mitochondrial large rRNA. This short sequence is sufficient to restore pole cell determination in UV-irradiated Drosophila embryos. Here, we provide evidence that the conserved 15-base sequence is sufficient to restore luciferase activity in vitro. Moreover, we show that the internal GAGA sequence is involved in protein binding and that mutations in this tetranucleotide affect the sequence’s ability to restore luciferase activity. The obtained results lead us to propose that mtlrRNA may be involved either in damaged protein reactivation or in protein biosynthesis during pole cell determination.
[1] Davidson, E.H. Developmental biology at the systems level. Biochem. Biophys. Acta 1789 (2009) 248–249. Search in Google Scholar
[2] Beams, H.W. and Kessel, R.G. The problem of germ cell determinants. Int. Rev. Cytol. 39 (1974) 413–479. http://dx.doi.org/10.1016/S0074-7696(08)60944-410.1016/S0074-7696(08)60944-4Search in Google Scholar
[3] Eddy, E.M. Germ plasm and the differentiation of the germ cell line. Int. Rev. Cytol. 43 (1975) 229–280. http://dx.doi.org/10.1016/S0074-7696(08)60070-410.1016/S0074-7696(08)60070-4Search in Google Scholar
[4] Ephrussi, A. and Lehmann, R. Induction of germ cell formation by oskar. Nature 358 (1992) 387–392. http://dx.doi.org/10.1038/358387a010.1038/358387a0Search in Google Scholar
[5] Nakamura, A., Amikura, R., Mukai, M., Kobayashi, S. and Lasko, P.F. Requirement for a noncoding RNA in Drosophila polar granules for germ cell establishment. Science 274 (1996) 2075–2079. http://dx.doi.org/10.1126/science.274.5295.207510.1126/science.274.5295.2075Search in Google Scholar
[6] Breitwieser, W., Markussen, F.H., Horstmann, H. and Ephrussi, A. Oskar protein interaction with Vasa represents an essential step in polar granules assembly. Genes Dev. 10 (1996) 2179–2188. http://dx.doi.org/10.1101/gad.10.17.217910.1101/gad.10.17.2179Search in Google Scholar
[7] Amikura, R., Kashikawa, M., Nakamura, A. and Kobayashi, S. Presence of mitochondria-type ribosomes outside mitochondria in germ plasm of Drosophila embryos. Proc. Natl. Acad. Sci. USA 98 (2001) 9133–9138. http://dx.doi.org/10.1073/pnas.17128699810.1073/pnas.171286998Search in Google Scholar
[8] Okada, M. Germline cell formation in Drosophila embryogenesis. Genes Genet. Syst. 73 (1998) 1–8. http://dx.doi.org/10.1266/ggs.73.110.1266/ggs.73.1Search in Google Scholar
[9] Kobayashi, S., Amikura, R. and Okada, M. Presence of mitochondrial large ribosomal RNA outside mitochondria in germ plasm of Drosophila melanogaster. Science 260 (1993) 1521–1524. http://dx.doi.org/10.1126/science.768485710.1126/science.7684857Search in Google Scholar
[10] Kobayashi, S. and Okada, M. Restoration of pole-cell-forming ability to u.v.-irradiated. Development 107 (1989) 733–42. Search in Google Scholar
[11] Kobayashi, S. and Okada, M. Complete cDNA sequence encoding mitochondrial large ribosomal RNA of Drosophila melanogaster. Nucleic Acids Res. 18 (1990) 4592. http://dx.doi.org/10.1093/nar/18.15.459210.1093/nar/18.15.4592Search in Google Scholar
[12] Jongens, T.A., Hay, B., Jan, L.Y. and Jan, Y.N. The germ cell-less gene product: a posteriorly localized component necessary for cell development in Drosophila. Cell 70 (1992) 569–584. http://dx.doi.org/10.1016/0092-8674(92)90427-E10.1016/0092-8674(92)90427-ESearch in Google Scholar
[13] Amikura, R., Sato, K. and Kobayashi, S. Role of mitochondrial ribosomedependent translation in germline formation in Drosophila embryos. Mech. Develop. 122 (2005) 1087–1093. http://dx.doi.org/10.1016/j.mod.2005.06.00310.1016/j.mod.2005.06.003Search in Google Scholar
[14] Braig, H.R., Guzman, H., Tesh R.B. and O’Neil, S.L. Replacement of the natural Wolbachia symbiont of Drosophila simulans with a mosquito counterpart. Nature 367 (1994) 453–455. http://dx.doi.org/10.1038/367453a010.1038/367453a0Search in Google Scholar
[15] Sambrook, J., Fritsch, E. and Maniatis, T. Molecular cloning. A Laboratory Manual. 2nd Edition. New York: Cold Spring Harbor Laboratory Press, 1989. Search in Google Scholar
[16] Ban, N., Nissen, P., Hansen, J., Moore, P.B. and Steitz, T.A. The complete atomic structure of the large ribosomal subunit at 2.4 A resolution. Science 289 (2000) 905–920. http://dx.doi.org/10.1126/science.289.5481.90510.1126/science.289.5481.905Search in Google Scholar
[17] Schluenzen, F., Tocilj, A., Zarivach, R., Harms, J., Gluehmann, M., Janell, D., Bashan, A., Bartels, H., Agmon, I., Franceschi, F. and Yonath, A. Structure of functionally activated small ribosomal subunit at 3.3 angstroms resolution. Cell 102 (2000) 615–623. http://dx.doi.org/10.1016/S0092-8674(00)00084-210.1016/S0092-8674(00)00084-2Search in Google Scholar
[18] Moazed, D. and Noller, H.F. Interaction of antibiotics with functional sites in 16S ribosomal RNA. Nature 327 (1987) 389–394. http://dx.doi.org/10.1038/327389a010.1038/327389a0Search in Google Scholar
[19] Nissen, P., Hansen, J., Ban, N., Moore, P.B. and Steitz, T.A. The structural basis of ribosome activity in peptide bond synthesis. Science 289 (2000) 920–930. http://dx.doi.org/10.1126/science.289.5481.92010.1126/science.289.5481.920Search in Google Scholar
[20] Hausner, T.P., Atmadja, J. and Nierhaus, K.H. Evidence that the G2661 region of 23S rRNA is located at the ribosomal binding sites of both elongation factors. Biochimie 69 (1987) 911–923. http://dx.doi.org/10.1016/0300-9084(87)90225-210.1016/0300-9084(87)90225-2Search in Google Scholar
[21] Moazed, D., Robertson, J.M. and Noller, H.F. Interaction of elongation factors EF-G and EF-Tu with a conserved loop in 23S RNA. Nature 334 (1988) 362–364. http://dx.doi.org/10.1038/334362a010.1038/334362a0Search in Google Scholar PubMed
[22] Emelyanov, V.V. Rickettsiaceae, Rickettsia-like endosymbionts, and the origin of mitochondria. Biosci. Rep. 21 (2001) 1–17. http://dx.doi.org/10.1023/A:101040941572310.1023/A:1010409415723Search in Google Scholar
[23] Emelyanov, V.V. Mitochondrial connection to the origin of the eukaryotic cell. Eur. J. Biochem. 270 (2003) 1599–1618. http://dx.doi.org/10.1046/j.1432-1033.2003.03499.x10.1046/j.1432-1033.2003.03499.xSearch in Google Scholar PubMed
[24] Chattopadhyay, S., Das, B. and DasGupta, C. Reactivation of denatured proteins by 23S ribosomal RNA: role of domain V. Proc. Natl. Acad. Sci. USA 93 (1996) 8284–8287. http://dx.doi.org/10.1073/pnas.93.16.828410.1073/pnas.93.16.8284Search in Google Scholar
[25] Grossweiner, L.I. Photochemical inactivation of enzymes. Curr. Top. Radiat. Res. Q. 11 (1976) 141–199. Search in Google Scholar
[26] Davies, M.J. Singlet oxygen-mediated damage to proteins and its consequences. Biochem. Biophys. Res. Commun. 305 (2003) 761–770. http://dx.doi.org/10.1016/S0006-291X(03)00817-910.1016/S0006-291X(03)00817-9Search in Google Scholar
[27] Pal, S., Chandra, S., Chowdhury, S., Sarkar, D., Ghosh, A.N. and DasGupta, C. Complementary role of two fragments of domain V of 23S ribosomal RNA in protein folding. J. Biol. Chem. 274 (1999) 32771–32777. http://dx.doi.org/10.1074/jbc.274.46.3277110.1074/jbc.274.46.32771Search in Google Scholar PubMed
[28] Sulijoadikusumo, I., Horikoshi, N. and Usheva, A. Another function for the mitochondrial ribosomal RNA: protein folding. Biochemistry 40 (2001) 11559–11564. http://dx.doi.org/10.1021/bi015526q10.1021/bi015526qSearch in Google Scholar PubMed
[29] Sanyal, S.C., Pal, S., Chowdhury, S., DasGupta, C. and Chowdhury, S. 23S rRNA assisted folding of cytoplasmic malate dehydrogenase is distinctly different from its self-folding. Nucleic Acids Res. 30 (2002) 2390–2397. http://dx.doi.org/10.1093/nar/30.11.239010.1093/nar/30.11.2390Search in Google Scholar PubMed PubMed Central
[30] Chowdhury, S., Pal, S., Ghosh, J. and DasGupta, C. Mutations in domain V of the 23S ribosomal RNA of Bacillus subtilis that inactivate its protein folding property in vitro. Nucleic Acids Res. 30 (2002) 1278–1285. http://dx.doi.org/10.1093/nar/30.5.127810.1093/nar/30.5.1278Search in Google Scholar PubMed PubMed Central
[31] Mahowald, A.P., Illmensee, K. and Turner, F.R. Interspecific transplantation of polar plasm between Drosophila embryos. J. Cell. Biol. 70 (1976) 358–373. http://dx.doi.org/10.1083/jcb.70.2.35810.1083/jcb.70.2.358Search in Google Scholar PubMed PubMed Central
[32] Illmensee, K., Mahowald, A.P. and Loomis, M.R. The ontogeny of germ plasm. Development 109 (1976) 425–33. Search in Google Scholar
© 2010 University of Wrocław, Poland
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.