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. 2013 Dec;163(4):1623-39.
doi: 10.1104/pp.113.223925. Epub 2013 Sep 13.

The REIL1 and REIL2 proteins of Arabidopsis thaliana are required for leaf growth in the cold

Stefanie Schmidt  1 , Frederik Dethloff, Olga Beine-Golovchuk, Joachim Kopka
Affiliations

The REIL1 and REIL2 proteins of Arabidopsis thaliana are required for leaf growth in the cold

Stefanie Schmidt et al. Plant Physiol. 2013 Dec.

Abstract

The evolutionarily conserved proteins REI1-LIKE (REIL1) and REIL2 have four conserved zinc finger domains and are Arabidopsis thaliana homologs of the cytosolic 60S ribosomal maturation factor Rei1p (for Required for isotropic bud growth1 protein) from yeast (Saccharomyces cerevisiae) and its paralog Reh1p (for REI1 homologue1 protein). The yeast and A. thaliana paralogs result from independent gene duplications. The A. thaliana REIL paralogs are required specifically in the cold (10°C) but not for growth at optimal temperature (20°C). A reil1-1 reil2-1 double mutant is arrested at 10°C prior to the emergence of the first rosette leaf. Two allelic reil2 mutants, reil2-1 and reil2-2, form small spoon-shaped leaves at 10°C. This phenomenon reverts after emergence of the inflorescence in the cold or upon shift to 20°C. Except for a slightly delayed germination, a reil1-1 mutant shows no further growth phenotype under the currently investigated conditions. A comparative analysis demonstrates conserved coexpression of orthologous genes from yeast and A. thaliana that are coregulated with yeast rei1 or with A. thaliana REIL2, respectively. The conserved correlations point to a role of A. thaliana REIL proteins in the maturation of the eukaryotic ribosomal 60S subunit. We support this conclusion by heterologous complementation of the cold-induced growth defect of the yeast Δrei1 deletion.

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Figures

Figure 1.
Figure 1.
Topology and phylogeny of the REIL proteins from A. thaliana. A, Topology of A. thaliana REIL genes and proteins. The REIL1 and REIL2 genes have five conserved exons. The respective REIL proteins, namely the 404-amino acid REIL1 and the 395-amino acid REIL2 protein, contain four zinc finger domains and two additional conserved domains, CD1 and CD2. The mutants, reil1-1 (SALK_090486), reil2-1 (GK_166C10), and reil2-2 (SALK_040068), carry T-DNA insertions in exon 2 (compare with arrow heads). B, Phylogenetic analysis of the plant REIL proteins. The plant REIL proteins were subject to gene duplications, which occurred independently in several plant phylae. The REIL1 and the REIL2 paralogs of A. thaliana originated from gene duplication during the speciation of the Brassicales.
Figure 2.
Figure 2.
The germination phenotype of reil mutants. A, Germination of reil mutants in the cold (10°C) compared with the Col-0 wild type. B, Germination of reil mutants at optimal temperature (20°C) compared with the Col-0 wild type. Significant differences relative to the wild type are indicated within the top section of each graph by circles, squares, diamonds, and triangles (P < 0.001, n = 4–5 plates per genotype). Each plate had approximately 80 seeds. The percentage of germinated seeds with cotyledons (left) and of seeds with a first rosette leaf greater than 1 mm (right) was scored per plate. The total number of germinated seeds per plate (100%) was scored at the last time point of each assay. Note that the rosette leaves of the reil1-1 reil2-1 double mutant emerged later at optimal temperature and did not emerge in the cold (arrow).
Figure 3.
Figure 3.
The aberrant leaf phenotypes of the reil1-1 reil2-1 double mutant and of the reil2-1 mutant. A, Pointed-leaves morphology of the reil1-1 reil2-1 double mutant compared with Col-0 after 16 d at 20°C. B, Growth arrest of the reil1-1 reil2-1 double mutant at stage 1.0 (Boyes et al., 2001) after 27 d at 10°C compared with Col-0. C, Phenotype of the reil1-1 reil2-1 double mutant germinated at 10°C, transferred to soil, and kept for 10 weeks strictly at 10°C compared with the Col-0 wild type. D, Representative phenotype of the reil2-1 mutant germinated at 10°C, transferred to soil, and kept for 5 to 6 weeks strictly at 10°C compared with the Col-0 wild type.
Figure 4.
Figure 4.
Phenotype of the reil1-1, reil2-1, and reil2-2 mutants compared with the Col-0 wild type under diverse temperature regimes. A, Phenotype after germination at 10°C, transfer to soil at stage 1.02 to 1.03, and continued growth at 10°C for a period of 4 weeks. B, Phenotype after germination at 10°C, transfer to soil at stage 1.02 to 1.03, and continued growth at 10°C for a period of 8 weeks. Note reversal of the reil2 leaf phenotype and emergence of inflorescences from all mutant and wild-type rosettes. C, Phenotype after germination at 10°C, transfer to soil at stage 1.02 to 1.03, continued growth at 10°C for a period of 2 weeks, and subsequent shift to 2 weeks at 20°C. Note reversal of the reil2 leaf phenotype after shift to optimum temperature. All photographs have equal scale. Vertically aligned photographs were taken with identical illumination.
Figure 5.
Figure 5.
Morphometric analyses of the reil1-1, reil2-1, and reil2-2 mutants compared with the Col-0 wild type. A, Plants cultivated strictly at 10°C (compare with Fig. 4, A and B). B, Plants germinated at 10°C, transferred to soil at stage 1.02 to 1.03, kept at 10°C for 2 weeks, and shifted to 20°C (compare with arrow and Fig. 4C). Significant differences relative to the wild type are indicated by circles, squares, and diamonds within the top section of each graph (P < 0.01, n = 5–10).
Figure 6.
Figure 6.
Mapping of A. thaliana orthologs to the scheme of the currently known cytosolic maturation machinery of the 60S large ribosomal subunit (Lo et al., 2010). Note that the ortholog of nmd3 was robustly coexpressed with REIL2 both under cold and under temperature control (black box). The other boxed orthologs were robustly coexpressed with REIL2 either under cold control or under temperature control. An asterisk indicates yeast genes without currently known A. thaliana orthologs.
Figure 7.
Figure 7.
Comparison of the coexpression matrix of the yeast 60S cytosolic maturation machinery to the coexpression matrix of A. thaliana paralogs. A, Yeast coexpression matrix under temperature control. B, A. thaliana coexpression matrix under temperature control. Note that average Spearman correlation coefficients (r) were calculated using independent transcript profiling experiments. The color scale represents positive coexpression with a maximum of +1.0 coded by red, with a minimal coexpression of –1.0 coded by blue and with white representing 0.0, i.e. the absence of coexpression. Empty cells were introduced in cases of gene families and for missing transcriptome data. Thirty-nine percent of the correlation coefficients match between A. thaliana and yeast with a deviation of r less than 0.250.
Figure 8.
Figure 8.
Complementation of the growth phenotype of the yeast Δrei1 mutant by heterologous expression of REIL1 or REIL2. The growth phenotype was characterized by calculations of the maximum relative growth rate (µmax) and the lag phase (λ) from semilogarithmic plots of growth curves (compare with Supplemental Fig. S11). The complementation is compared at suboptimal temperatures, 28°C and 22°C, to the BY4742 wild type (wt, black bars), the yeast Δrei1 mutant (–, white bars), and the yeast Δrei1 mutant complemented by homologous expression of yeast rei1 (Rei1p, dark gray bars). Note that the white squares at the top of each graph indicate significant differences compared with the Δrei1 mutant, i.e. complementation. Black squares indicate fully or partially maintained significant differences compared with the wild type. Three independently transformed strains expressing REIL1 or REIL2 were tested (P < 0.01, n = 16–32).
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References

    1. Agarwal P, Arora R, Ray S, Singh AK, Singh VP, Takatsuji H, Kapoor S, Tyagi AK. (2007) Genome-wide identification of C2H2 zinc-finger gene family in rice and their phylogeny and expression analysis. Plant Mol Biol 65: 467–485 - PubMed
    1. Alberti S, Gitler AD, Lindquist S. (2007) A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae. Yeast 24: 913–919 - PMC - PubMed
    1. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402 - PMC - PubMed
    1. Basu U, Si K, Warner JR, Maitra U. (2001) The Saccharomyces cerevisiae TIF6 gene encoding translation initiation factor 6 is required for 60S ribosomal subunit biogenesis. Mol Cell Biol 21: 1453–1462 - PMC - PubMed
    1. Berná G, Robles P, Micol JL. (1999) A mutational analysis of leaf morphogenesis in Arabidopsis thaliana. Genetics 152: 729–742 - PMC - PubMed

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