Abstract
The high-chlorophyll fluorescence photosynthesis mutant hcf109 of Arabidopsis was characterized in detail to gain insights into the regulatory mechanism of RNA processing in higher plants. By using electron transport, chlorophyll fluorescence, and immunoblot studies, we assigned the mutational lesion to photosystems I and II and the plastid NAD(P)H dehydrogenase complex. The functional pleiotropy was reflected in RNA deficiencies. Although all nuclear-encoded photosynthetic RNAs analyzed revealed no difference in size or steady state level between mutant and wild type, the RNA patterns of the plastome-encoded psbB-psbT-psbH-petB-petD, psbD-psbC-ycf9, ndhC-ndhK-ndhJ, and ndhH-ndhA-ndhI-ndhG-ndhE-psaC-ndh D transcription units were severely disturbed. These operons encode subunits of photosystems I (psa) and II (psb), the cytochrome bGf complex (pet), the plastid NAD(P)H dehydrogenase (ndh), and the unidentified open reading frame ycf9. With the exception of the ndhC operon, the RNA deficiencies observed were specific and restricted to particular segments of the psbB, psbD/C, and ndhH operons, that is, the psbB-psbT, ycf9, and psaC regions. Run-on transcription studies with isolated chloroplasts showed that the failure of these transcripts to accumulate was due to RNA stability and not transcription. Other polycistronic transcription units analyzed were not affected by the mutation. This result indicates that the trans-regulatory factor encoded by the hcf109 gene is not a general RNA stability factor but that it specifically controls the stability of only these distinct transcripts. Because the hcf109 locus was mapped at a distance < 0.1 centimorgans from the phytochrome C gene, its molecular characterization by positional cloning is possible.
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- Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barkan A., Miles D., Taylor W. C. Chloroplast gene expression in nuclear, photosynthetic mutants of maize. EMBO J. 1986 Jul;5(7):1421–1427. doi: 10.1002/j.1460-2075.1986.tb04378.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barkan A. Proteins encoded by a complex chloroplast transcription unit are each translated from both monocistronic and polycistronic mRNAs. EMBO J. 1988 Sep;7(9):2637–2644. doi: 10.1002/j.1460-2075.1988.tb03116.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barkan A., Walker M., Nolasco M., Johnson D. A nuclear mutation in maize blocks the processing and translation of several chloroplast mRNAs and provides evidence for the differential translation of alternative mRNA forms. EMBO J. 1994 Jul 1;13(13):3170–3181. doi: 10.1002/j.1460-2075.1994.tb06616.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bell C. J., Ecker J. R. Assignment of 30 microsatellite loci to the linkage map of Arabidopsis. Genomics. 1994 Jan 1;19(1):137–144. doi: 10.1006/geno.1994.1023. [DOI] [PubMed] [Google Scholar]
- Berends T., Gamble P. E., Mullet J. E. Characterization of the barley chloroplast transcription units containing psaA-psaB and psbD-psbC. Nucleic Acids Res. 1987 Jul 10;15(13):5217–5240. doi: 10.1093/nar/15.13.5217. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blowers A. D., Klein U., Ellmore G. S., Bogorad L. Functional in vivo analyses of the 3' flanking sequences of the Chlamydomonas chloroplast rbcL and psaB genes. Mol Gen Genet. 1993 Apr;238(3):339–349. doi: 10.1007/BF00291992. [DOI] [PubMed] [Google Scholar]
- Båga M., Göransson M., Normark S., Uhlin B. E. Processed mRNA with differential stability in the regulation of E. coli pilin gene expression. Cell. 1988 Jan 29;52(2):197–206. doi: 10.1016/0092-8674(88)90508-9. [DOI] [PubMed] [Google Scholar]
- Chen H. C., Stern D. B. Specific ribonuclease activities in spinach chloroplasts promote mRNA maturation and degradation. J Biol Chem. 1991 Dec 15;266(35):24205–24211. [PubMed] [Google Scholar]
- Christopher D. A., Kim M., Mullet J. E. A novel light-regulated promoter is conserved in cereal and dicot chloroplasts. Plant Cell. 1992 Jul;4(7):785–798. doi: 10.1105/tpc.4.7.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dieckmann C. L., Koerner T. J., Tzagoloff A. Assembly of the mitochondrial membrane system. CBP1, a yeast nuclear gene involved in 5' end processing of cytochrome b pre-mRNA. J Biol Chem. 1984 Apr 25;259(8):4722–4731. [PubMed] [Google Scholar]
- Eddy C. K., Keshav K. F., An H., Utt E. A., Mejia J. P., Ingram L. O. Segmental message stabilization as a mechanism for differential expression from the Zymomonas mobilis gap operon. J Bacteriol. 1991 Jan;173(1):245–254. doi: 10.1128/jb.173.1.245-254.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gamble P. E., Sexton T. B., Mullet J. E. Light-dependent changes in psbD and psbC transcripts of barley chloroplasts: accumulation of two transcripts maintains psbD and psbC translation capability in mature chloroplasts. EMBO J. 1988 May;7(5):1289–1297. doi: 10.1002/j.1460-2075.1988.tb02943.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grivell L. A. Nucleo-mitochondrial interactions in mitochondrial gene expression. Crit Rev Biochem Mol Biol. 1995;30(2):121–164. doi: 10.3109/10409239509085141. [DOI] [PubMed] [Google Scholar]
- Haley J., Bogorad L. Alternative promoters are used for genes within maize chloroplast polycistronic transcription units. Plant Cell. 1990 Apr;2(4):323–333. doi: 10.1105/tpc.2.4.323. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hallick R. B., Lipper C., Richards O. C., Rutter W. J. Isolation of a transcriptionally active chromosome from chloroplasts of Euglena gracilis. Biochemistry. 1976 Jul 13;15(14):3039–3045. doi: 10.1021/bi00659a016. [DOI] [PubMed] [Google Scholar]
- Hudson G. S., Mason J. G., Holton T. A., Koller B., Cox G. B., Whitfeld P. R., Bottomley W. A gene cluster in the spinach and pea chloroplast genomes encoding one CF1 and three CF0 subunits of the H+-ATP synthase complex and the ribosomal protein S2. J Mol Biol. 1987 Jul 20;196(2):283–298. doi: 10.1016/0022-2836(87)90690-5. [DOI] [PubMed] [Google Scholar]
- Jensen K. H., Herrin D. L., Plumley F. G., Schmidt G. W. Biogenesis of photosystem II complexes: transcriptional, translational, and posttranslational regulation. J Cell Biol. 1986 Oct;103(4):1315–1325. doi: 10.1083/jcb.103.4.1315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kapoor S., Wakasugi T., Deno H., Sugiura M. An atpE-specific promoter within the coding region of the atpB gene in tobacco chloroplast DNA. Curr Genet. 1994 Sep;26(3):263–268. doi: 10.1007/BF00309558. [DOI] [PubMed] [Google Scholar]
- Klein R. R., Mullet J. E. Light-induced transcription of chloroplast genes. psbA transcription is differentially enhanced in illuminated barley. J Biol Chem. 1990 Feb 5;265(4):1895–1902. [PubMed] [Google Scholar]
- Klimyuk V. I., Carroll B. J., Thomas C. M., Jones J. D. Alkali treatment for rapid preparation of plant material for reliable PCR analysis. Plant J. 1993 Mar;3(3):493–494. doi: 10.1111/j.1365-313x.1993.tb00169.x. [DOI] [PubMed] [Google Scholar]
- Konieczny A., Ausubel F. M. A procedure for mapping Arabidopsis mutations using co-dominant ecotype-specific PCR-based markers. Plant J. 1993 Aug;4(2):403–410. doi: 10.1046/j.1365-313x.1993.04020403.x. [DOI] [PubMed] [Google Scholar]
- Kuchka M. R., Goldschmidt-Clermont M., van Dillewijn J., Rochaix J. D. Mutation at the Chlamydomonas nuclear NAC2 locus specifically affects stability of the chloroplast psbD transcript encoding polypeptide D2 of PS II. Cell. 1989 Sep 8;58(5):869–876. doi: 10.1016/0092-8674(89)90939-2. [DOI] [PubMed] [Google Scholar]
- Lander E. S., Green P., Abrahamson J., Barlow A., Daly M. J., Lincoln S. E., Newberg L. A., Newburg L. MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics. 1987 Oct;1(2):174–181. doi: 10.1016/0888-7543(87)90010-3. [DOI] [PubMed] [Google Scholar]
- Matsubayashi T., Wakasugi T., Shinozaki K., Yamaguchi-Shinozaki K., Zaita N., Hidaka T., Meng B. Y., Ohto C., Tanaka M., Kato A. Six chloroplast genes (ndhA-F) homologous to human mitochondrial genes encoding components of the respiratory chain NADH dehydrogenase are actively expressed: determination of the splice sites in ndhA and ndhB pre-mRNAs. Mol Gen Genet. 1987 Dec;210(3):385–393. doi: 10.1007/BF00327187. [DOI] [PubMed] [Google Scholar]
- McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meurer J., Meierhoff K., Westhoff P. Isolation of high-chlorophyll-fluorescence mutants of Arabidopsis thaliana and their characterisation by spectroscopy, immunoblotting and northern hybridisation. Planta. 1996;198(3):385–396. doi: 10.1007/BF00620055. [DOI] [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]
- Monod C., Takahashi Y., Goldschmidt-Clermont M., Rochaix J. D. The chloroplast ycf8 open reading frame encodes a photosystem II polypeptide which maintains photosynthetic activity under adverse growth conditions. EMBO J. 1994 Jun 15;13(12):2747–2754. doi: 10.1002/j.1460-2075.1994.tb06568.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newbury S. F., Smith N. H., Higgins C. F. Differential mRNA stability controls relative gene expression within a polycistronic operon. Cell. 1987 Dec 24;51(6):1131–1143. doi: 10.1016/0092-8674(87)90599-x. [DOI] [PubMed] [Google Scholar]
- Nickelsen J., Link G. The 54 kDa RNA-binding protein from mustard chloroplasts mediates endonucleolytic transcript 3' end formation in vitro. Plant J. 1993 Apr;3(4):537–544. doi: 10.1046/j.1365-313x.1993.03040537.x. [DOI] [PubMed] [Google Scholar]
- Nickelsen J., van Dillewijn J., Rahire M., Rochaix J. D. Determinants for stability of the chloroplast psbD RNA are located within its short leader region in Chlamydomonas reinhardtii. EMBO J. 1994 Jul 1;13(13):3182–3191. doi: 10.1002/j.1460-2075.1994.tb06617.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poetsch W., Hermans J., Westhoff P. Multiple cDNAs of phosphoenolpyruvate carboxylase in the C4 dicot Flaveria trinervia. FEBS Lett. 1991 Nov 4;292(1-2):133–136. doi: 10.1016/0014-5793(91)80850-3. [DOI] [PubMed] [Google Scholar]
- Reinbothe S., Reinbothe C., Heintzen C., Seidenbecher C., Parthier B. A methyl jasmonate-induced shift in the length of the 5' untranslated region impairs translation of the plastid rbcL transcript in barley. EMBO J. 1993 Apr;12(4):1505–1512. doi: 10.1002/j.1460-2075.1993.tb05794.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]
- Sakamoto W., Kindle K. L., Stern D. B. In vivo analysis of Chlamydomonas chloroplast petD gene expression using stable transformation of beta-glucuronidase translational fusions. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):497–501. doi: 10.1073/pnas.90.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
- Sieburth L. E., Berry-Lowe S., Schmidt G. W. Chloroplast RNA Stability in Chlamydomonas: Rapid Degradation of psbB and psbC Transcripts in Two Nuclear Mutants. Plant Cell. 1991 Feb;3(2):175–189. doi: 10.1105/tpc.3.2.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stern D. B., Gruissem W. Control of plastid gene expression: 3' inverted repeats act as mRNA processing and stabilizing elements, but do not terminate transcription. Cell. 1987 Dec 24;51(6):1145–1157. doi: 10.1016/0092-8674(87)90600-3. [DOI] [PubMed] [Google Scholar]
- Stern D. B., Radwanski E. R., Kindle K. L. A 3' stem/loop structure of the Chlamydomonas chloroplast atpB gene regulates mRNA accumulation in vivo. Plant Cell. 1991 Mar;3(3):285–297. doi: 10.1105/tpc.3.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Woodbury N. W., Dobres M., Thompson W. F. The identification and localization of 33 pea chloroplast transcription initiation sites. Curr Genet. 1989 Dec;16(5-6):433–445. doi: 10.1007/BF00340723. [DOI] [PubMed] [Google Scholar]
- von Gabain A., Belasco J. G., Schottel J. L., Chang A. C., Cohen S. N. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]