Abstract
Most of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes characterized in plants and algae to date have one intron very close to the 5′ end of the gene. To study the functional relevance of some of these introns for gene expression we have analysed the influence of three 5′ introns on transient gene expression of the anaerobically inducible maizeGapC4 promoter in maize cells. Under aerobic conditions, reporter gene expression is increased in the presence of the first introns of theGapC4 andGapC1 genes, and the first intron of the nuclear encoded chloroplast-specificGapA1 gene. In contrast, theGapC4 intron increases anaerobic gene expression above the level obtained for the intronless construct, while anaerobic expression of constructs harboring theGapA1 andGapC1 introns was similar to the anaerobic expression level of the intronless construct. Splicing analysis revealed that theGapC4 intron is processed more efficiently under anaerobic conditions, while no change in splicing efficiency is observed for theGapC1 and theGapA1 introns when subjected to anaerobic conditions. These results suggest that an increase in splicing efficiency contributes to the anaerobic induction of the maizeGapC4 gene.
References
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 7:248–254
Callis J, Fromm M, Walbot V (1987) Introns increase gene expression in cultured maize cells. Genes Dev 1:1183–1200
Carle-Urioste J, Ko CH, Benito M-I, Walbot V (1994) In vivo analysis of intron processing using splicing-dependent reporter gene assays. Plant Mol Biol 26:1785–1795
Cerff R (1995) The chimeric nature of nuclear genomes and the antiquity of introns as demonstrated by the GAPDH gene system. In: Go M, Schimmel P (eds) Proceedings of the 20th Taniguchi International Symposium “Tracing biological evolution in protein and gene structures”, Nagoya, Japan. Elsevier Science, Amsterdam, pp 203–225
Czarnecka E, Nagao RT, Key JL Gurley WB (1988) Characterization ofGmhsp26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition of intron processing. Mol Cell Biol 8:1113–1122
DeWet JR, Wood KV, DeLuca M, Helinski DR, Subramani S (1987) Firefly luciferase gene: structure and expression in mammalian cells. Mol Cell Biol 7:725–737
Donath M, Mendel R, Cerff R, Martin W (1995) Intron-dependent transient expression of the maizeGapA1 gene. Plant Mol Biol 28:667–676
Goodall GJ, Filipowicz W (1991) Different effects of intron nucleotide composition and secondary structure on pre-mRNA splicing in monocot and dicot plants. EMBO J 10:2635–2644
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusuions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Kersanach R, Brinkmann H, Liaud M-F, Zhang D-X, Martin WF, Cerff R (1994) Five identical intron positions in ancient duplicated genes of eubacterial origin. Nature 367: 387–389
Klein TM, Gradziel T, Fromm ME, Sanford JC (1988) Factors influencing gene delivery intoZea mays cells by high-velocity microprojectiles. Bio/Tech 6:559–563
Köhler U, Liaud M-F, Mendel RR, Cerff R, Hehl R (1995) The maizeGapC4 promoter confers anaerobic reporter gene expression and shows homology to the maize anthocyanin regulatory locusC1. Plant Mol Biol, 29:1293–1298
Last DI, Brettel RIS, Chamberlain DA, Chaudhury AM, Larkin PJ, Marsh EL, Peacock WJ, Dennis ES (1991) pEmu: an improved vector for gene expression in cereal cells. Theor Appl Genet 81:581–588
Lévesque G, Bharucha AD, Murthy MRV (1993) Reverse transcription and PCR amplification of rare mRNAs immobilized on oligo(dT) cellulose. Anal Biochem 213:170–171
Liaud MF, Brandt U, Cerff R (1995) The marine red algaChondrus crispus has a highly divergent beta-tubulin gene with a characteristic 5′ intron: functional and evolutionary implications. Plant Mol Biol 28:313–325
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissue. Anal Biochem 163:16–20
Luehrsen KR, Walbot V (1991) Intron enhancement of gene expression and the splicing efficiency of introns in maize cells. Mol Gen Genet 225:81–93
Maas C, Laufs J, Grant S, Korfhage C, Werr W (1991) The combination of a novel stimulatory element in the first exon of the maizeShrunkenI gene with the following intron 1 enhances reporter gene expression up to 1000-fold. Plant Mol Biol 16:199–207
Martinez P, Martin W, Cerff R (1989) Structure, evolution and anaerobic regulation of a nuclear gene encoding cytosolic glycer-aldehyde-3-phosphate dehydrogenase from maize. J Mol Biol 208:551–565
Mascarenhas D, Mettler IJ, Pierce DA, Lowe HW (1990) Intronmediated enhancement of heterologous gene expression in maize. Plant Mol Biol 15:913–920
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497
Nash J, Walbot V (1992)Bronze-2 gene expression and intron splicing patterns in cells and tissues ofZea mays L. Plant Physiol 100:464–471
Norris SR, Meyer SE, Callis J (1993) The intron of aArabidopsis thaliana polyubiquitin gene is conserved in location and is quantitative determinant of chimaeric gene expression. Plant Mol Biol 21:895–906
Olson DC, Oetiker JH, Yang SF (1995) Analysis ofLE-ACS3, a 1-aminocyclopropane-1-carboxylic acid synthase gene expressed during flooding in the roots of tomato plants. J Biol Chem 270:14056–14061
Ortiz DF, Strommer JN (1990) TheMul maize transposable element induces tissue-specific aberrant splicing and polyadenylation in twoAdh1 mutants. Mol Cell Biol 10:2090–2095
Osteryoung KW, Sundberg H, Vierling E (1993) Poly(A) tail length of a heat shock protein RNA is increased by severe heat stress, but intron splicing is unaffected. Mol Gen Genet 239:323–333
Quigley F, Martin WF, Cerff R (1988) Intron conservation across the prokaryote-eukaryote boundary: structure of the nuclear gene for chloroplast glyceraldehyde-3-phosphate dehydrogenase from maize. Proc Natl Acad Sci USA 85:2672–2676
Rathus C, Bower R, Birch RG (1993) Effects of promoter, intron, and enhancer elements on transient gene expression in sugarcane and carrot protoplasts. Plant Mol Biol 23:613–618
Rochester DE, Winter JA, Shah DM (1986) The structure and expression of maize genes encoding the major heat shock protein, hsp 70. EMBO J 5:451–458
Russell DA, Sachs MM (1989) Differential expression and sequence analysis of the maize glyceraldehyde-3-phosphate dehydrogenase gene family. Plant Cell 1:793–803
Russell DA, Sachs MM (1992) Protein synthesis in maize during anaerobic and heat stress. Plant Physiol 99:615–620
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual: (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sanford JC (1988) The biolistic process. Trends Biotech 6:299–302
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain termination inhibitors. Proc Nat Acad Sci USA 74:5463–5467
Schledzewski K, Mendel RR (1994) Quantitative transient gene expression: comparison of the promoters for maize polyubiquitin1, rice actin1, maize-derived Emu and CaMV 35S in cells of bareley, maize and tobacco. Transgenic Res 3:249–255
Sinibaldi RM, Mettler IJ (1992) Intron splicing and intron-mediated enhanced expression in monocots. In: Progress in nucleic acid research and molecular biology. Vol. 42. Academic Press, pp 229–257
Tanaka A, Mita S, Ohta S, Kyozuka J, Shimamoto K, Nakamura K (1990) Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron. Nucleic Acids Res 18:6767–6770
Töpfer R, Pröls M, Schell J, Steinbiss HH (1988) Transient gene expression in tobacco protoplasts: comparison of reporter gene systems for CAT, NPTII, and GUS. Plant Cell Rep 7:225–228
Vain P, McCullen MD, Finer JJ (1993) Osmotic treatment enhances particle bombardment-mediated transient and stable transformation of maize. Plant Cell Rep 12:84–88
Winter J, Wright R, Duck N, Gasser C, Fraley R, Shah D (1988) The inhibition of petuniahsp70 mRNA processing during CdCl2 stress. Mol Gen Genet 211:315–319
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Köhler, U., Cerff, R., Hehl, R. et al. Intron-specific stimulation of anaerobic gene expression and splicing efficiency in maize cells. Molec. Gen. Genet. 251, 252–258 (1996). https://doi.org/10.1007/BF02172925
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DOI: https://doi.org/10.1007/BF02172925