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Homodimerization of HYL1 ensures the correct selection of cleavage sites in primary miRNA.

Yang X, Ren W, Zhao Q, Zhang P, Wu F, He Y - Nucleic Acids Res. (2014)

Bottom Line: Disruption of HYL1 homodimerization causes incorrect cleavage at sites in pri-miRNA without interrupting the interaction of HYL1 with DCL1 and accumulation of pri-miRNAs in HYL1/pri-miRNA complexes, leading to a reduction in the efficiency and accuracy of miRNAs that results in strong mutant phenotypes of the plants.HYL1 homodimers may function as a molecular anchor for DCL1 to cleave at a distance from the ssRNA-dsRNA junction in pri-miRNA.These results suggest that HYL1 ensures the correct selection of pri-miRNA cleavage sites through homodimerization and thus contributes to gene silencing and plant development.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

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Accumulation of some miRNAs and expression of their targets in the plants deficient in HYL1 homodimerization. (A) Northern blot showing miRNA accumulation in the hyl1-2 mutants and the transgenic plants. (B) The expression levels of some miRNA-targeted genes in the hyl1-2 mutants and the transgenic plants. (C) Detection of HYL1 homodimers in the hyl1-3 mutants. HYL1 antibody was used to detect HYL1 from the total protein extract of the hyl1-3 plants using the buffer without addition of SDS and urea.
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Figure 5: Accumulation of some miRNAs and expression of their targets in the plants deficient in HYL1 homodimerization. (A) Northern blot showing miRNA accumulation in the hyl1-2 mutants and the transgenic plants. (B) The expression levels of some miRNA-targeted genes in the hyl1-2 mutants and the transgenic plants. (C) Detection of HYL1 homodimers in the hyl1-3 mutants. HYL1 antibody was used to detect HYL1 from the total protein extract of the hyl1-3 plants using the buffer without addition of SDS and urea.

Mentions: To examine whether the deficiency in HYL1 dimerization affects miRNA biogenesis, we performed northern blotting to detect the accumulation of miRNAs in the hyl1-2 mutants and transgenic plants with HYL1 mutations. In the hyl1-2;G147E, hyl1-2;L165E plants, the accumulation of miR156, miR166, miR172 and miR319 was much less than that in wild-type plants (Figure 5A). By contrast, miR156-targeted SPL9 and miR165/6-targeted REV genes were up-regulated in these plants (Figure 5B). Moreover, the accumulation of these miRNAs in hyl1-2;T146E, hyl1-2;I158E and hyl1-2;L166E was much greater than that in hyl1-2, whereas the target genes SPL9 and REV were down-regulated to the levels almost equal to those of the wild-type.


Homodimerization of HYL1 ensures the correct selection of cleavage sites in primary miRNA.

Yang X, Ren W, Zhao Q, Zhang P, Wu F, He Y - Nucleic Acids Res. (2014)

Accumulation of some miRNAs and expression of their targets in the plants deficient in HYL1 homodimerization. (A) Northern blot showing miRNA accumulation in the hyl1-2 mutants and the transgenic plants. (B) The expression levels of some miRNA-targeted genes in the hyl1-2 mutants and the transgenic plants. (C) Detection of HYL1 homodimers in the hyl1-3 mutants. HYL1 antibody was used to detect HYL1 from the total protein extract of the hyl1-3 plants using the buffer without addition of SDS and urea.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
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Figure 5: Accumulation of some miRNAs and expression of their targets in the plants deficient in HYL1 homodimerization. (A) Northern blot showing miRNA accumulation in the hyl1-2 mutants and the transgenic plants. (B) The expression levels of some miRNA-targeted genes in the hyl1-2 mutants and the transgenic plants. (C) Detection of HYL1 homodimers in the hyl1-3 mutants. HYL1 antibody was used to detect HYL1 from the total protein extract of the hyl1-3 plants using the buffer without addition of SDS and urea.
Mentions: To examine whether the deficiency in HYL1 dimerization affects miRNA biogenesis, we performed northern blotting to detect the accumulation of miRNAs in the hyl1-2 mutants and transgenic plants with HYL1 mutations. In the hyl1-2;G147E, hyl1-2;L165E plants, the accumulation of miR156, miR166, miR172 and miR319 was much less than that in wild-type plants (Figure 5A). By contrast, miR156-targeted SPL9 and miR165/6-targeted REV genes were up-regulated in these plants (Figure 5B). Moreover, the accumulation of these miRNAs in hyl1-2;T146E, hyl1-2;I158E and hyl1-2;L166E was much greater than that in hyl1-2, whereas the target genes SPL9 and REV were down-regulated to the levels almost equal to those of the wild-type.

Bottom Line: Disruption of HYL1 homodimerization causes incorrect cleavage at sites in pri-miRNA without interrupting the interaction of HYL1 with DCL1 and accumulation of pri-miRNAs in HYL1/pri-miRNA complexes, leading to a reduction in the efficiency and accuracy of miRNAs that results in strong mutant phenotypes of the plants.HYL1 homodimers may function as a molecular anchor for DCL1 to cleave at a distance from the ssRNA-dsRNA junction in pri-miRNA.These results suggest that HYL1 ensures the correct selection of pri-miRNA cleavage sites through homodimerization and thus contributes to gene silencing and plant development.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.

Show MeSH