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PMID:15821876

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Citation

Hiraguri, A, Itoh, R, Kondo, N, Nomura, Y, Aizawa, D, Murai, Y, Koiwa, H, Seki, M, Shinozaki, K and Fukuhara, T (2005) Specific interactions between Dicer-like proteins and HYL1/DRB-family dsRNA-binding proteins in Arabidopsis thaliana. Plant Mol. Biol. 57:173-88

Abstract

Proteins that specifically bind double-stranded RNA (dsRNA) are involved in the regulation of cellular signaling events and gene expression, and are characterized by a conserved dsRNA-binding motif (dsRBM). Here we report the biochemical properties of nine such gene products, each containing one or two dsRBMs: four Arabidopsis Dicer-like proteins (DCL1-4), Arabidopsis HYL1 and four of its homologs (DRB2, DRB4, DRB5 and OsDRB1). DCL1, DCL3, HYL1 and the four HYL1 homologs exhibit significant dsRNA-binding activity, indicating that these proteins are involved in RNA metabolism. The dsRBMs from dsRBM-containing proteins (dsRBPs) also function as a protein-protein interaction domain and homo- and heterodimerization are essential for biological functioning of these proteins. We show that DRB4 interacts specifically with DCL4, and HYL1 most strongly interacts with DCL1. These results indicate that each HYL1/DRB family protein interacts with one specific partner among the four Dicer-like proteins. Localization studies using GFP fusion proteins demonstrate that DCL1, DCL4, HYL1 and DRB4 localize in the nucleus, while DRB2 is present in the cytoplasm. Subcellular localizations of HYL1, DRB4, DCL1 and DCL4 further strengthen the notion that HYL1 and DCL1, and DRB4 and DCL4, exist as complexes. The presented data suggest that each member of the HYL1/DRB protein family may individually modulate Dicer function through heterodimerization with a Dicer-like protein in vivo.

Links

PubMed Online version:10.1007/s11103-004-6853-5

Keywords

Arabidopsis/genetics; Arabidopsis/metabolism; Arabidopsis Proteins/chemistry; Arabidopsis Proteins/genetics; Arabidopsis Proteins/metabolism; Binding, Competitive; Cell Cycle Proteins/chemistry; Cell Cycle Proteins/genetics; Cell Cycle Proteins/metabolism; Dimerization; Electrophoresis, Polyacrylamide Gel; Electrophoretic Mobility Shift Assay; Green Fluorescent Proteins/genetics; Green Fluorescent Proteins/metabolism; Microscopy, Fluorescence; Oryza sativa/genetics; Oryza sativa/metabolism; Phylogeny; Plant Proteins/genetics; Plant Proteins/metabolism; Protein Binding; RNA, Double-Stranded/metabolism; RNA-Binding Proteins/chemistry; RNA-Binding Proteins/genetics; RNA-Binding Proteins/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism; Ribonuclease III/chemistry; Ribonuclease III/genetics; Ribonuclease III/metabolism; Tradescantia/cytology; Tradescantia/genetics; Tradescantia/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ARATH:F4HQG6

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:F4HQG6

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

AGI_LocusCode:AT1G09700

F

Seeded From UniProt

complete

ARATH:F4HQG6

part_of

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8H1D4

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8GY79

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SKN2

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

AGI_LocusCode:AT1G01040

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0042802: identical protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SP32

F

Seeded From UniProt

complete

ARATH:DRB1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q3EBC8

F

Seeded From UniProt

complete

ARATH:DCL4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8H1D4

F

Seeded From UniProt

complete

ARATH:DCL4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

AGI_LocusCode:AT3G62800

F

Seeded From UniProt

complete

ORYSJ:DRB8

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:DCL2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9LXW7

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SKN2

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SP32

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8H1D4

F

Seeded From UniProt

complete

ARATH:DRB5

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:DRB4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8GY79

F

Seeded From UniProt

complete

ARATH:DRB4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DRB4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P84634

F

Seeded From UniProt

complete

ARATH:DRB4

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

AGI_LocusCode:AT5G20320

F

Seeded From UniProt

complete

ARATH:DCL3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SKN2

F

Seeded From UniProt

complete

ARATH:DCL3

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8GY79

F

Seeded From UniProt

complete

ARATH:DCL3

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:DRB2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9LXW7

F

Seeded From UniProt

complete

ARATH:DRB2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SP32

F

Seeded From UniProt

complete

ARATH:DRB2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8GY79

F

Seeded From UniProt

complete

ARATH:DRB2

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DRB2

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

ARATH:DRB2

part_of

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:DCL1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q8GY79

F

Seeded From UniProt

complete

ARATH:DCL1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q9SKN2

F

Seeded From UniProt

complete

ARATH:DCL1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:O04492

F

Seeded From UniProt

complete

ARATH:DCL4

part_of

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:DRB4

part_of

GO:0005634: nucleus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

ARATH:DRB4

enables

GO:0003725: double-stranded RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

See also

References

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