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PMID:17110334

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Citation

Yigit, E, Batista, PJ, Bei, Y, Pang, KM, Chen, CC, Tolia, NH, Joshua-Tor, L, Mitani, S, Simard, MJ and Mello, CC (2006) Analysis of the C. elegans Argonaute family reveals that distinct Argonautes act sequentially during RNAi. Cell 127:747-57

Abstract

Argonaute (AGO) proteins interact with small RNAs to mediate gene silencing. C. elegans contains 27 AGO genes, raising the question of what roles these genes play in RNAi and related gene-silencing pathways. Here we describe 31 deletion alleles representing all of the previously uncharacterized AGO genes. Analysis of single- and multiple-AGO mutant strains reveals functions in several pathways, including (1) chromosome segregation, (2) fertility, and (3) at least two separate steps in the RNAi pathway. We show that RDE-1 interacts with trigger-derived sense and antisense RNAs to initiate RNAi, while several other AGO proteins interact with amplified siRNAs to mediate downstream silencing. Overexpression of downstream AGOs enhances silencing, suggesting that these proteins are limiting for RNAi. Interestingly, these AGO proteins lack key residues required for mRNA cleavage. Our findings support a two-step model for RNAi, in which functionally and structurally distinct AGOs act sequentially to direct gene silencing.

Links

PubMed Online version:10.1016/j.cell.2006.09.033

Keywords

Amino Acid Sequence; Animals; Caenorhabditis elegans/metabolism; Caenorhabditis elegans Proteins/chemistry; Caenorhabditis elegans Proteins/metabolism; Embryo, Nonmammalian/cytology; Models, Genetic; Molecular Sequence Data; Mutation/genetics; Phylogeny; Protein Binding; RNA Interference; RNA, Helminth/metabolism; RNA, Small Interfering/metabolism; Recombinant Fusion Proteins/metabolism

Significance

Annotations

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

CAEEL:G5EEH0

enables

GO:0035197: siRNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

CAEEL:G5EEH0

part_of

GO:0016442: RISC complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

CAEEL:P90786

involved_in

GO:0000003: reproduction

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:O61931

involved_in

GO:0016246: RNA interference

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:Q9GZG4

GO:1900369: negative regulation of RNA interference

IMP: Inferred from Mutant Phenotype:

P

Figure 4A

complete
CACAO 7444


CAEEL:Q9GZG4

GO:1900369: negative regulation of RNA interference

IMP: Inferred from Mutant Phenotype:

P

Seeded From UniProt

complete


CAEEL:H2KZD5

involved_in

GO:0016246: RNA interference

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:H2KZD5

involved_in

GO:0000070: mitotic sister chromatid segregation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

CAEEL:ERGO1

involved_in

GO:0016246: RNA interference

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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