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PMID:20504953

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Citation

Temme, C, Zhang, L, Kremmer, E, Ihling, C, Chartier, A, Sinz, A, Simonelig, M and Wahle, E (2010) Subunits of the Drosophila CCR4-NOT complex and their roles in mRNA deadenylation. RNA 16:1356-70

Abstract

The CCR4-NOT complex is the main enzyme catalyzing the deadenylation of mRNA. We have investigated the composition of this complex in Drosophila melanogaster by immunoprecipitation with a monoclonal antibody directed against NOT1. The CCR4, CAF1 (=POP2), NOT1, NOT2, NOT3, and CAF40 subunits were associated in a stable complex, but NOT4 was not. Factors known to be involved in mRNA regulation were prominent among the other proteins coprecipitated with the CCR4-NOT complex, as analyzed by mass spectrometry. The complex was localized mostly in the cytoplasm but did not appear to be a major component of P bodies. Of the known CCR4 paralogs, Nocturnin was found associated with the subunits of the CCR4-NOT complex, whereas Angel and 3635 were not. RNAi experiments in Schneider cells showed that CAF1, NOT1, NOT2, and NOT3 are required for bulk poly(A) shortening and hsp70 mRNA deadenylation, but knock-down of CCR4, CAF40, and NOT4 did not affect these processes. Overexpression of catalytically dead CAF1 had a dominant-negative effect on mRNA decay. In contrast, overexpression of inactive CCR4 had no effect. We conclude that CAF1 is the major catalytically important subunit of the CCR4-NOT complex in Drosophila Schneider cells. Nocturnin may also be involved in mRNA deadenylation, whereas there is no evidence for a similar role of Angel and 3635.

Links

PubMed PMC2885685 Online version:10.1261/rna.2145110

Keywords

Adenosine Monophosphate/metabolism; Animals; Cell Line; Drosophila Proteins/analysis; Drosophila Proteins/metabolism; Drosophila melanogaster/enzymology; Drosophila melanogaster/metabolism; Proteome/analysis; RNA, Messenger/metabolism; Retinoblastoma-Binding Protein 4/metabolism; Ribonucleases/chemistry; Ribonucleases/metabolism

Significance

Annotations

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

DROME:A0A0B4LEZ3

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:A0A0B4LEZ3

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:A0A0B4LEZ3

involved_in

GO:0000289: nuclear-transcribed mRNA poly(A) tail shortening

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q7JVP2

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q7K126

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q7K126

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q7K126

involved_in

GO:0000289: nuclear-transcribed mRNA poly(A) tail shortening

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q8IMX1

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q8MR41

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q8MR41

enables

GO:0004535: poly(A)-specific ribonuclease activity

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

DROME:RGA

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:RGA

located_in

GO:0005829: cytosol

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:RGA

involved_in

GO:0000289: nuclear-transcribed mRNA poly(A) tail shortening

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:CNO10

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:X2JCC3

part_of

GO:0030014: CCR4-NOT complex

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:X2JCC3

involved_in

GO:0000289: nuclear-transcribed mRNA poly(A) tail shortening

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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