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PMID:11344258

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Citation

Gross, S and Moore, C (2001) Five subunits are required for reconstitution of the cleavage and polyadenylation activities of Saccharomyces cerevisiae cleavage factor I. Proc. Natl. Acad. Sci. U.S.A. 98:6080-5

Abstract

Cleavage and polyadenylation of mRNA 3' ends in Saccharomyces cerevisiae requires several factors, one of which is cleavage factor I (CF I). Purification of CF I activity from yeast extract has implicated numerous proteins as functioning in both cleavage and/or polyadenylation. Through reconstitution of active CF I from separately expressed and purified proteins, we show that CF I contains five subunits, Rna14, Rna15, Pcf11, Clp1, and Hrp1. These five are necessary and sufficient for reconstitution of cleavage activity in vitro when mixed with CF II, and for specific polyadenylation when mixed with polyadenylation factor I, purified poly(A) polymerase, and poly(A) binding protein. Analysis of the individual protein-protein interactions supports an architectural model for CF I in which Pcf11 simultaneously interacts with Rna14, Rna15, and Clp1, whereas Rna14 bridges Rna15 and Hrp1.

Links

PubMed PMC33425 Online version:10.1073/pnas.101046598

Keywords

Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/isolation & purification; Adenosine Triphosphatases/metabolism; Binding Sites; DNA Helicases/genetics; DNA Helicases/isolation & purification; DNA Helicases/metabolism; Fungal Proteins/genetics; Fungal Proteins/isolation & purification; Fungal Proteins/metabolism; Gene Expression; Nuclear Proteins/genetics; Nuclear Proteins/isolation & purification; Nuclear Proteins/metabolism; Poly A/metabolism; RNA-Binding Proteins/genetics; RNA-Binding Proteins/isolation & purification; RNA-Binding Proteins/metabolism; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/isolation & purification; Recombinant Fusion Proteins/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism; Saccharomyces cerevisiae Proteins; mRNA Cleavage and Polyadenylation Factors

Significance

Annotations

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

YEAST:RNA14

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:RNA14

part_of

GO:0005849: mRNA cleavage factor complex

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25299,UniProtKB:P39081,UniProtKB:Q08685

C

Seeded From UniProt

complete

YEAST:RNA14

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:RNA14

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25299

F

Seeded From UniProt

complete

YEAST:RNA14

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P39081

F

Seeded From UniProt

complete

YEAST:RNA14

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q99383

F

Seeded From UniProt

complete

YEAST:RNA15

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:RNA15

part_of

GO:0005849: mRNA cleavage factor complex

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25298,UniProtKB:P39081,UniProtKB:Q08685

C

Seeded From UniProt

complete

YEAST:RNA15

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:RNA15

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25298

F

Seeded From UniProt

complete

YEAST:RNA15

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P39081

F

Seeded From UniProt

complete

YEAST:PCF11

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PCF11

part_of

GO:0005849: mRNA cleavage factor complex

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q08685

C

Seeded From UniProt

complete

YEAST:PCF11

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PCF11

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25298

F

Seeded From UniProt

complete

YEAST:PCF11

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25299

F

Seeded From UniProt

complete

YEAST:PCF11

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:Q08685

F

Seeded From UniProt

complete

YEAST:PFS2

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:PFS2

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:FIP1

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:FIP1

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:YTH1

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:YTH1

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CFT1

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CFT1

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CLP1

involved_in

GO:0031124: mRNA 3'-end processing

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CLP1

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CLP1

contributes_to

GO:0003723: RNA binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

YEAST:CLP1

part_of

GO:0005849: mRNA cleavage factor complex

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P39081

C

Seeded From UniProt

complete

YEAST:CLP1

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CLP1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P39081

F

Seeded From UniProt

complete

YEAST:CFT2

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:CFT2

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:HRP1

involved_in

GO:0006378: mRNA polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:HRP1

part_of

GO:0005849: mRNA cleavage factor complex

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25298,UniProtKB:P25299,UniProtKB:P39081,UniProtKB:Q08685

C

Seeded From UniProt

complete

YEAST:HRP1

involved_in

GO:0098789: pre-mRNA cleavage required for polyadenylation

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:HRP1

enables

GO:0005515: protein binding

ECO:0000353: physical interaction evidence used in manual assertion

UniProtKB:P25298

F

Seeded From UniProt

complete


See also

References

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