Molecular evolution of duplicate copies of genes encoding cytosolic glutamine synthetase in Pisum sativum |
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Authors: | Elsbeth L. Walker N. F. Weeden Crispin B. Taylor Pamela Green Gloria M. Coruzzi |
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Affiliation: | (1) Dept. of Biology, New York University, 1009 Main Bldg, 10003 New York, NY, USA;(2) Department of Horticultural Science, New York State Agricultural experiment Station, Box 462, 14456 Geneva, NY, USA;(3) MSU-DOE Plant Research Laboratory, Michigan State University, 48824-1312 East Lansing, MI, USA;(4) Department of Biochemistry, Michigan State University, 48824-1312 East Lansing, MI, USA;(5) Present address: Department of Biological Sciences, Mount Holyoke College, So. Hadley, 01075, MA, USA;(6) Present address: Dept. of Biology, University of North Carolina, 27599-3280 Chapel Hill, NC, USA |
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Abstract: | Here, we describe two nearly identical expressed genes for cytosolic glutamine synthetase (GS3A and GS3B) in Pisum sativum L. RFLP mapping data indicates that the GS3A and GS3B genes are separate loci located on different chromosomes. DNA sequencing of the GS3A and GS3B genes revealed that the coding regions are 99% identical with only simple nucleotide substitutions resulting in three amino acid differences. Surprisingly, the non-coding regions (5 non-coding leader, the 11 introns, and 3 non-coding tail) all showed a high degree of identity (96%). In these non-coding regions, 25% of the observed differences between the GS3A and GS3B genes were deletions or duplications. The single difference in the 3 non-coding regions of the GS3A and GS3B genes was a 25 bp duplication of an AU-rich element in the GS3B gene. As the GS3B mRNA accumulates to lower levels than the GS3A gene, we tested whether this sequence which resembles an mRNA instability determinant functioned as such in the context of the GS mRNA. Using the GS3B 3 tail as part of a chimeric gene in transgenic plants, we showed that this AU-rich sequence has little effect on transgene mRNA levels. To determine whether the GS3A/GS3B genes represent a recent duplication, we examined GS3-like genes in genomic DNA of ancient relatives of P. sativum. We observed that several members of the Viceae each contain two genomic DNA fragments homologous to the GS3B gene, suggesting that this is an ancient duplication event. Gene conversion has been invoked as a possible mechanism for maintaining the high level of nucleotide similarity found between the GS3A and GS3B genes. Possible evolutionary reasons for the maintenance of these twin GS genes in pea, and the general duplication of genes for cytosolic GS in all plant species are discussed. |
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Keywords: | Glutamine synthetase gene duplication multigene family molecular evolution gene conversion |
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