Structure of arabidopsis Hyponastic Leaves1 and its molecular implications for miRNA Processing

Seong Wook Yang; Hong Ying Chen; Jing Yang; Satoru Machida; Nam Hai Chua; Y. Adam Yuan

doi:10.1016/j.str.2010.02.006

Structure of arabidopsis Hyponastic Leaves1 and its molecular implications for miRNA Processing

Seong Wook Yang, Hong Ying Chen, Jing Yang, Satoru Machida, Nam Hai Chua, Y. Adam Yuan

Research output: Contribution to journal › Article › peer-review

122 Citations (Scopus)

Abstract

The Arabidopsis HYPONASTIC LEAVES1 (HYL1) is a double-stranded RNA-binding protein that forms a complex with DICER-LIKE1 (DCL1) and SERRATE to facilitate processing of primary miRNAs into microRNAs (miRNAs). However, the structural mechanisms of miRNA maturation by this complex are poorly understood. Here, we present the crystal structures of double-stranded RNA binding domains (dsRBD1 and dsRBD2) of HYL1 and HYL1 dsRBD1 (HR1)/dsRNA complex as well as human TRBP2 dsRBD2 (TR2)/dsRNA complex for comparison analysis. Structural and functional study demonstrates that both HR1 and TR2 are canonical dsRBDs for dsRNA binding, whereas HR2 of HYL1 is a non-canonical dsRBD harboring a putative dimerization interface. Domain swapping within the context of HYL1 demonstrates that TR2 can supplant the function of HR1 in vitro and in vivo. Further biochemical analyses suggest that HYL1 probably binds to the miRNA/miRNA^* region of precursors as a dimer mediated by HR2.

Original language	English
Pages (from-to)	594-605
Number of pages	12
Journal	Structure
Volume	18
Issue number	5
DOIs	https://doi.org/10.1016/j.str.2010.02.006
Publication status	Published - 2010 May

Bibliographical note

Funding Information:
We thank A. Saxena at X12C at the National Synchrotron Light Source of the Brookhaven National Laboratory for assistance with data collection. This work was supported by intramural research funds from Temasek Life Sciences Laboratory, National University of Singapore Research Scholarship, and Singapore Ministry of Education (T208A3124 to Y.A.Y.). Work performed at the Rockefeller University was supported by National Institutes of Health grant GM44640 to N.-H.C. S.W.Y. performed biochemical and genetic analyses; H.-Y.C. performed constructs screening, subcloning, and protein crystallization; J.Y. performed ITC and gel filtration analysis; S.M. performed HYL1 dimer detection; and Y.A.Y. solved the structures. Y.A.Y., S.W.Y., and N.-H.C. wrote the paper with input from H.-Y.C.

All Science Journal Classification (ASJC) codes

Structural Biology
Molecular Biology

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10.1016/j.str.2010.02.006

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title = "Structure of arabidopsis Hyponastic Leaves1 and its molecular implications for miRNA Processing",

abstract = "The Arabidopsis HYPONASTIC LEAVES1 (HYL1) is a double-stranded RNA-binding protein that forms a complex with DICER-LIKE1 (DCL1) and SERRATE to facilitate processing of primary miRNAs into microRNAs (miRNAs). However, the structural mechanisms of miRNA maturation by this complex are poorly understood. Here, we present the crystal structures of double-stranded RNA binding domains (dsRBD1 and dsRBD2) of HYL1 and HYL1 dsRBD1 (HR1)/dsRNA complex as well as human TRBP2 dsRBD2 (TR2)/dsRNA complex for comparison analysis. Structural and functional study demonstrates that both HR1 and TR2 are canonical dsRBDs for dsRNA binding, whereas HR2 of HYL1 is a non-canonical dsRBD harboring a putative dimerization interface. Domain swapping within the context of HYL1 demonstrates that TR2 can supplant the function of HR1 in vitro and in vivo. Further biochemical analyses suggest that HYL1 probably binds to the miRNA/miRNA* region of precursors as a dimer mediated by HR2.",

author = "Yang, {Seong Wook} and Chen, {Hong Ying} and Jing Yang and Satoru Machida and Chua, {Nam Hai} and Yuan, {Y. Adam}",

note = "Funding Information: We thank A. Saxena at X12C at the National Synchrotron Light Source of the Brookhaven National Laboratory for assistance with data collection. This work was supported by intramural research funds from Temasek Life Sciences Laboratory, National University of Singapore Research Scholarship, and Singapore Ministry of Education (T208A3124 to Y.A.Y.). Work performed at the Rockefeller University was supported by National Institutes of Health grant GM44640 to N.-H.C. S.W.Y. performed biochemical and genetic analyses; H.-Y.C. performed constructs screening, subcloning, and protein crystallization; J.Y. performed ITC and gel filtration analysis; S.M. performed HYL1 dimer detection; and Y.A.Y. solved the structures. Y.A.Y., S.W.Y., and N.-H.C. wrote the paper with input from H.-Y.C. ",

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AU - Yuan, Y. Adam

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Structure of arabidopsis Hyponastic Leaves1 and its molecular implications for miRNA Processing

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

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