Abstract
Translation of the chloroplast psbC mRNA in Chlamydomonas reinhardtii has been shown previously to require interactions between its 5' untranslated region (5' UTR) and the functions encoded by two nuclear loci, which we name here TBC1 and TBC2. We show that a 97-nucleotide (nt) region located in the middle of the psbC 5' UTR is required for translation initiation. Unlike most procaryotic cis-acting translational control elements, this region has a translational activation function and is located 236 nt upstream from the GUG translation initiation codon. In vivo pulse-labeling of chloroplast-encoded proteins and analyses of the expression of chimeric reporter genes in vivo reveal that a mutation of a newly described locus, TBC3, restores translation from the psbC 5' UTR in the absence of either this cis-acting element or the wild-type trans-acting TBC1 function. These data demonstrate that sequences located in the middle of the psbC 5' UTR, TBC1, and TBC3 functionally interact to control the translation of the psbC mRNA.
Full Text
The Full Text of this article is available as a PDF (1.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bennoun P., Masson A., Delosme M. A method for complementation analysis of nuclear and chloroplast mutants of photosynthesis in chlamydomonas. Genetics. 1980 May;95(1):39–47. doi: 10.1093/genetics/95.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Costanzo M. C., Fox T. D. Specific translational activation by nuclear gene products occurs in the 5' untranslated leader of a yeast mitochondrial mRNA. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2677–2681. doi: 10.1073/pnas.85.8.2677. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Costanzo M. C., Fox T. D. Suppression of a defect in the 5' untranslated leader of mitochondrial COX3 mRNA by a mutation affecting an mRNA-specific translational activator protein. Mol Cell Biol. 1993 Aug;13(8):4806–4813. doi: 10.1128/mcb.13.8.4806. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Danon A., Mayfield S. P. Light-regulated translation of chloroplast messenger RNAs through redox potential. Science. 1994 Dec 9;266(5191):1717–1719. doi: 10.1126/science.7992056. [DOI] [PubMed] [Google Scholar]
- Delepelaire P. Partial characterization of the biosynthesis and integration of the Photosystem II reaction centers in the thylakoid membrane of Chlamydomonas reinhardtii. EMBO J. 1984 Apr;3(4):701–706. doi: 10.1002/j.1460-2075.1984.tb01872.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gillham N. W., Boynton J. E., Hauser C. R. Translational regulation of gene expression in chloroplasts and mitochondria. Annu Rev Genet. 1994;28:71–93. doi: 10.1146/annurev.ge.28.120194.000443. [DOI] [PubMed] [Google Scholar]
- Girard-Bascou J., Pierre Y., Drapier D. A nuclear mutation affects the synthesis of the chloroplast psbA gene production Chlamydomonas reinhardtii. Curr Genet. 1992 Jul;22(1):47–52. doi: 10.1007/BF00351741. [DOI] [PubMed] [Google Scholar]
- Gold L. Posttranscriptional regulatory mechanisms in Escherichia coli. Annu Rev Biochem. 1988;57:199–233. doi: 10.1146/annurev.bi.57.070188.001215. [DOI] [PubMed] [Google Scholar]
- Goldschmidt-Clermont M., Girard-Bascou J., Choquet Y., Rochaix J. D. Trans-splicing mutants of Chlamydomonas reinhardtii. Mol Gen Genet. 1990 Sep;223(3):417–425. doi: 10.1007/BF00264448. [DOI] [PubMed] [Google Scholar]
- Goldschmidt-Clermont M. Transgenic expression of aminoglycoside adenine transferase in the chloroplast: a selectable marker of site-directed transformation of chlamydomonas. Nucleic Acids Res. 1991 Aug 11;19(15):4083–4089. doi: 10.1093/nar/19.15.4083. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gorman D. S., Levine R. P. Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci U S A. 1965 Dec;54(6):1665–1669. doi: 10.1073/pnas.54.6.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris E. H., Boynton J. E., Gillham N. W. Chloroplast ribosomes and protein synthesis. Microbiol Rev. 1994 Dec;58(4):700–754. doi: 10.1128/mr.58.4.700-754.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hirose T., Sugiura M. Cis-acting elements and trans-acting factors for accurate translation of chloroplast psbA mRNAs: development of an in vitro translation system from tobacco chloroplasts. EMBO J. 1996 Apr 1;15(7):1687–1695. [PMC free article] [PubMed] [Google Scholar]
- Hwang S., Kawazoe R., Herrin D. L. Transcription of tufA and other chloroplast-encoded genes is controlled by a circadian clock in Chlamydomonas. Proc Natl Acad Sci U S A. 1996 Feb 6;93(3):996–1000. doi: 10.1073/pnas.93.3.996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuchka M. R., Mayfield S. P., Rochaix J. D. Nuclear mutations specifically affect the synthesis and/or degradation of the chloroplast-encoded D2 polypeptide of photosystem II in Chlamydomonas reinhardtii. EMBO J. 1988 Feb;7(2):319–324. doi: 10.1002/j.1460-2075.1988.tb02815.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LEVINE R. P., EBERSOLD W. T. The genetics and cytology of Chlamydomonas. Annu Rev Microbiol. 1960;14:197–216. doi: 10.1146/annurev.mi.14.100160.001213. [DOI] [PubMed] [Google Scholar]
- Mayfield S. P., Cohen A., Danon A., Yohn C. B. Translation of the psbA mRNA of Chlamydomonas reinhardtii requires a structured RNA element contained within the 5' untranslated region. J Cell Biol. 1994 Dec;127(6 Pt 1):1537–1545. doi: 10.1083/jcb.127.6.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McMullin T. W., Fox T. D. COX3 mRNA-specific translational activator proteins are associated with the inner mitochondrial membrane in Saccharomyces cerevisiae. J Biol Chem. 1993 Jun 5;268(16):11737–11741. [PubMed] [Google Scholar]
- Mittelmeier T. M., Dieckmann C. L. In vivo analysis of sequences required for translation of cytochrome b transcripts in yeast mitochondria. Mol Cell Biol. 1995 Feb;15(2):780–789. doi: 10.1128/mcb.15.2.780. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Monod C., Goldschmidt-Clermont M., Rochaix J. D. Accumulation of chloroplast psbB RNA requires a nuclear factor in Chlamydomonas reinhardtii. Mol Gen Genet. 1992 Feb;231(3):449–459. doi: 10.1007/BF00292715. [DOI] [PubMed] [Google Scholar]
- Ooi B. G., Lukins H. B., Linnane A. W., Nagley P. Biogenesis of mitochondria: a mutation in the 5'-untranslated region of yeast mitochondrial oli1 mRNA leading to impairment in translation of subunit 9 of the mitochondrial ATPase complex. Nucleic Acids Res. 1987 Mar 11;15(5):1965–1977. doi: 10.1093/nar/15.5.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rochaix J. D., Kuchka M., Mayfield S., Schirmer-Rahire M., Girard-Bascou J., Bennoun P. Nuclear and chloroplast mutations affect the synthesis or stability of the chloroplast psbC gene product in Chlamydomonas reinhardtii. EMBO J. 1989 Apr;8(4):1013–1021. doi: 10.1002/j.1460-2075.1989.tb03468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rochaix J. D. Post-transcriptional steps in the expression of chloroplast genes. Annu Rev Cell Biol. 1992;8:1–28. doi: 10.1146/annurev.cb.08.110192.000245. [DOI] [PubMed] [Google Scholar]
- Rochaix J. D. Restriction endonuclease map of the chloroplast DNA of Chlamydomonas reinhardii. J Mol Biol. 1978 Dec 25;126(4):597–617. doi: 10.1016/0022-2836(78)90011-6. [DOI] [PubMed] [Google Scholar]
- Rodermel S., Haley J., Jiang C. Z., Tsai C. H., Bogorad L. A mechanism for intergenomic integration: abundance of ribulose bisphosphate carboxylase small-subunit protein influences the translation of the large-subunit mRNA. Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3881–3885. doi: 10.1073/pnas.93.9.3881. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rutherford A. W. Photosystem II, the water-splitting enzyme. Trends Biochem Sci. 1989 Jun;14(6):227–232. doi: 10.1016/0968-0004(89)90032-7. [DOI] [PubMed] [Google Scholar]
- Sakamoto W., Chen X., Kindle K. L., Stern D. B. Function of the Chlamydomonas reinhardtii petd 5' untranslated region in regulating the accumulation of subunit IV of the cytochrome b6/f complex. Plant J. 1994 Oct;6(4):503–512. doi: 10.1046/j.1365-313x.1994.6040503.x. [DOI] [PubMed] [Google Scholar]
- Schmidt G. W., Bartlett S. G., Grossman A. R., Cashmore A. R., Chua N. H. Biosynthetic pathways of two polypeptide subunits of the light-harvesting chlorophyll a/b protein complex. J Cell Biol. 1981 Nov;91(2 Pt 1):468–478. doi: 10.1083/jcb.91.2.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
- Staub J. M., Maliga P. Accumulation of D1 polypeptide in tobacco plastids is regulated via the untranslated region of the psbA mRNA. EMBO J. 1993 Feb;12(2):601–606. doi: 10.1002/j.1460-2075.1993.tb05692.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiesenberger G., Costanzo M. C., Fox T. D. Analysis of the Saccharomyces cerevisiae mitochondrial COX3 mRNA 5' untranslated leader: translational activation and mRNA processing. Mol Cell Biol. 1995 Jun;15(6):3291–3300. doi: 10.1128/mcb.15.6.3291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wu H. Y., Kuchka M. R. A nuclear suppressor overcomes defects in the synthesis of the chloroplast psbD gene product caused by mutations in two distinct nuclear genes of Chlamydomonas. Curr Genet. 1995 Feb;27(3):263–269. doi: 10.1007/BF00326159. [DOI] [PubMed] [Google Scholar]
- Zerges W., Rochaix J. D. The 5' leader of a chloroplast mRNA mediates the translational requirements for two nucleus-encoded functions in Chlamydomonas reinhardtii. Mol Cell Biol. 1994 Aug;14(8):5268–5277. doi: 10.1128/mcb.14.8.5268. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Vitry C., Olive J., Drapier D., Recouvreur M., Wollman F. A. Posttranslational events leading to the assembly of photosystem II protein complex: a study using photosynthesis mutants from Chlamydomonas reinhardtii. J Cell Biol. 1989 Sep;109(3):991–1006. doi: 10.1083/jcb.109.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]