Cleavage Stimulation Factor

2011 ◽  
Keyword(s):  
Cleavage Stimulation Factor

scholarly journals Elimination of Introns at the Drosophila suppressor-of-forked Locus by P-Element-Mediated Gene Conversion Shows That an RNA Lacking a Stop Codon is Dispensable

Genetics ◽  
1996 ◽  
Vol 143 (1) ◽  
pp. 345-351
Author(s):  
Carol J Williams ◽  
Kevin O'Hare
Keyword(s):  
X Chromosome ◽  
Stop Codon ◽  
Alternative Polyadenylation ◽  
Gene Replacement ◽  
P Element ◽  
Wild Type ◽  
Genomic Location ◽  
Chromosomal Breaks ◽  
White Locus ◽  
Cleavage Stimulation Factor

Abstract The suppressor of forked [su(f)] locus affects the phenotype of mutations caused by transposable element insertions at unlinked loci. It encodes a putative 84-kD protein with homology to two proteins involved in mRNA 3′ end processing; the product of the yeast RNA14 gene and the 77-kD subunit of human cleavage stimulation factor. Three su(f) mRNAs are produced by alternative polyadenylation. The 2. 6 and 2.9-kb mRNAs encode the same 84-kD protein while a 1.3-kb RNA, which terminates within the fourth intron, is unusual in having no stop codon. Using P-element-mediated gene replacement we have copied sequences from a transformation construct into the su(f) gene creating a su(f) allele at the normal genomic location that lacks the first five introns. This allele is viable and appears wild type for su(f) function, demonstrating that the 1.3-kb RNA and the sequences contained within the deleted introns are dispensable for su(f) function. Compared with studies on gene replacement at the white locus, chromosomal breaks at su(f) appear to be less efficiently repaired from ectopic sites, perhaps because of the location of su(f) at the euchromatin/heterochromatin boundary on the X chromosome.

The Xenopus laevis homologue of the 64-kDa subunit of cleavage stimulation factor

1996 ◽  
Vol 114 (3) ◽  
pp. 313-315 ◽  
Author(s):  
S Barbaux ◽  
L.T Seery ◽  
D.R Schoenberg ◽  
G.C Sellar ◽  
A.S Whitehead
Keyword(s):  
Xenopus Laevis ◽  
Cleavage Stimulation Factor

scholarly journals A temperature sensitive mutation in the CstF77 subunit of the polyadenylation complex reveals the critical function of mRNA 3' end formation for a robust heat stress response in plants

2021 ◽  
Author(s):  
Minsoo Kim ◽  
John D Swenson ◽  
Fionn McLoughlin ◽  
Elizabeth Vierling
Keyword(s):  
Heat Stress ◽  
Molecular Mechanisms ◽  
Dominant Role ◽  
Temperature Sensitive ◽  
Heat Sensitivity ◽  
Critical Function ◽  
Heat Stress Response ◽  
Transcript Processing ◽  
Cleavage Stimulation Factor ◽  
Inducible Genes

Background: Heat Shock Protein 101 (HSP101) in plants and orthologs in bacteria (Caseinolytic peptidase B, ClpB) and yeast (Hsp104) are essential for thermotolerance. To investigate molecular mechanisms of thermotolerance involving HSP101, we performed a suppressor screen in Arabidopsis thaliana of a semi-dominant, missense HSP101 allele, hot1-4 (A499T). Plants carrying the hot1-4 mutation are more heat-sensitive than an HSP101 null mutant (hot1-3), indicating the toxicity of hot1-4 allele. Results: We report that one suppressor (shot2, suppressor of hot1-4 2) has a temperature-sensitive, missense mutation (E170K) in the CstF77 (Cleavage stimulation factor 77) subunit of the polyadenylation complex, which is critical for 3' end maturation of pre-mRNA. RNA-Seq analysis of total RNA depleted of ribosomes reveals that heat treatment causes transcriptional readthrough events in shot2, specifically in highly heat-induced genes, including the toxic hot1-4 gene. In addition, failure of correct transcript processing leads to reduced accumulation of many HSP RNAs and proteins, suppressing heat sensitivity of the hot1-4 mutant, due to reduction of the toxic mutant HSP101 protein. Notably, the shot2 mutation makes plants more sensitive to heat stress in the HSP101 null (hot1-3) and wild-type backgrounds correlated with the reduced expression of other heat-inducible genes required for thermotolerance. Conclusions: Our study reveals the critical function of CstF77 for 3' end formation of mRNA during heat stress, as well as the dominant role of HSP101 in dictating the outcome of severe heat stress in plants.

A multicomponent complex is required for the AAUAAA-dependent cross-linking of a 64-kilodalton protein to polyadenylation substrates

1990 ◽  
Vol 10 (3) ◽  
pp. 1244-1248 ◽  
Author(s):  
J Wilusz ◽  
T Shenk ◽  
Y Takagaki ◽  
J L Manley
Keyword(s):  
Uv Light ◽  
Cross Linking ◽  
Cross Link ◽  
Multicomponent Complex ◽  
Specificity Factor ◽  
The Individual ◽  
64 Kda Protein ◽  
Rna Interaction ◽  
Cleavage Stimulation Factor

A 64-kilodalton (kDa) polypeptide that is cross-linked by UV light specifically to polyadenylation substrate RNAs containing a functional AAUAAA element has been identified previously. Fractionated HeLa nuclear components that can be combined to regenerate efficient and accurate polyadenylation in vitro have now been screened for the presence of the 64-kDa protein. None of the individual components contained an activity which could generate the 64-kDa species upon UV cross-linking in the presence of substrate RNA. It was necessary to mix two components, cleavage stimulation factor and specificity factor, to reconstitute 64-kDa protein-RNA cross-linking. The addition of cleavage factors to this mixture very efficiently reconstituted the AAUAAA-specific 64-kDa protein-RNA interaction. The 64-kDa protein, therefore, is present in highly purified, reconstituted polyadenylation reactions. However, it is necessary to form a multicomponent complex to efficiently cross-link the protein to a substrate RNA.

scholarly journals Two distinct forms of the 64,000 Mr protein of the cleavage stimulation factor are expressed in mouse male germ cells

1999 ◽  
Vol 96 (12) ◽  
pp. 6763-6768 ◽  
Author(s):  
A. M. Wallace ◽  
B. Dass ◽  
S. E. Ravnik ◽  
V. Tonk ◽  
N. A. Jenkins ◽  
...  
Keyword(s):  
Germ Cells ◽  
Male Germ Cells ◽  
Cleavage Stimulation Factor

Polymorphism 10034C>T Is Located in a Functional Cleavage Stimulation Factor (CstF) Binding Site of the Fibrinogen Gamma Gene and Influences the γ′/γA mRNA Ratio.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 543-543 ◽  
Author(s):  
Shirley Uitte de Willige ◽  
Inge M. Rietveld ◽  
Marieke C.H. de Visser ◽  
Hans L. Vos ◽  
Rogier M. Bertina
Keyword(s):  
Venous Thrombosis ◽  
Deep Venous Thrombosis ◽  
Binding Site ◽  
Point Mutations ◽  
Fold Increase ◽  
Nucleotide Polymorphisms ◽  
Functional Variation ◽  
Increased Risk ◽  
Cleavage Stimulation Factor

Abstract Previously we found that haplotype 2 of the fibrinogen gamma gene (FGG-H2) is associated with an increased risk of deep venous thrombosis and with reduced fibrinogen γ′ levels, an alternatively spliced form of FGG, and reduced fibrinogen γ′/total fibrinogen ratios (Blood2005; 106(13): 4176–83). Two of the FGG-H2 tagging single nucleotide polymorphisms (SNPs), 9615C>T [rs2066864] and 10034C>T [rs2066865], are located in the region containing the two polyadenylation (pA) signals used in FGG pre-mRNA processing, namely pA1 and pA2, resulting in transcripts coding for γ′-chains and γA-chains, respectively. SNP 10034C>T is located in a Cleavage stimulation Factor (CstF) binding site at nucleotides 10030–10047 of FGG. CstF is a multi-subunit complex required for efficient cleavage and polyadenylation of pre-mRNAs. We hypothesized that 10034T, by strengthening this CstF binding site, favors use of pA2 (synthesis of γA-chains) at the expense of pA1-use (synthesis of γ′-chains). We transiently transfected FGG mini-gene constructs containing exon 9, intron 9 (containing pA1), exon 10 and the 3′ UTR (containing pA2) in liver-derived HepG2 cells and used quantitative real-time PCR to measure the mean relative pA-signal usage (pA1/pA2 ratio) of the different constructs. To study the effects of SNPs 9615C>T and 10034C>T on the relative use of pA1 and pA2, we made constructs carrying the different alleles of these SNPs. Compared to reference construct CC (9615C, 10034C; FGG-H1) of which the pA1/pA2 ratio was set at 100%, the ratio of construct TT (9615T, 10034T; FGG-H2) was 1.4-fold decreased (71.5%, p=0.015). To investigate whether the SNPs individually or collectively caused the reduction in the pA1/pA2 ratio, exchange constructs CT (9615C, 10034T) and TC (9615T, 10034C) were also tested. The ratio of construct CT was almost 1.2-fold decreased (85.3%, p=0.001), while the ratio of construct TC did not differ significantly from the reference construct (101.6%, p=0.890). These results indicate that in our in vitro system SNP 10034C>T, and not SNP 9615C>T, is the major contributor to the reduction of the pA1/pA2 ratio. We further investigated the functionality and importance of the CstF binding site, containing 10034C>T, with a construct in which nucleotides 10013–10056 containing the complete CstF binding site were deleted. Compared to the reference construct, in which the CstF site was still present, the pA1/pA2 ratio of the CstF deletion construct was 2.2-fold increased (222.3%, p<0.001), indicating that this site is involved in the regulation of pA2-usage. The functionality of the CstF site was further confirmed using several mutant constructs in which the CstF consensus was weakened or strengthened by introducing one or two point mutations. Weakening the CstF site resulted in a significant 1.5-fold increase of the pA1/pA2 ratio, while strengthening the site resulted in a significant 1.6-fold decrease of the ratio. From these results we conclude that 10034C>T is located in a functional CstF binding site and is involved in regulating the usage of the pA2-signal of FGG. We propose that the 10034C>T change is the functional variation in the FGG-H2 haplotype and is responsible for the reduction in the fibrinogen γ′/total fibrinogen ratio and the increased risk of deep venous thrombosis.

scholarly journals Characterization of a Cleavage Stimulation Factor, 3′ pre-RNA, Subunit 2, 64 kDa (CSTF2) as a Therapeutic Target for Lung Cancer

2011 ◽  
Vol 17 (18) ◽  
pp. 5889-5900 ◽  
Author(s):  
Masato Aragaki ◽  
Koji Takahashi ◽  
Hirohiko Akiyama ◽  
Eiju Tsuchiya ◽  
Satoshi Kondo ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Therapeutic Target ◽  
Cleavage Stimulation Factor
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