THO/Sub2p functions to coordinate 3'-end processing with gene-nuclear pore association.

During transcription, proteins assemble sequentially with nascent RNA to generate a messenger ribonucleoprotein particle (mRNP). The THO complex and its associated Sub2p helicase are functionally implicated in both transcription and mRNP biogenesis but their precise function remains elusive. We show here that THO/Sub2p mutation leads to the accumulation of a stalled intermediate in mRNP biogenesis that contains nuclear pore components and polyadenylation factors in association with chromatin. Microarray analyses of genomic loci that are aberrantly docked to the nuclear pore in mutants allowed the identification of approximately 400 novel validated target genes that require THO /Sub2p for efficient expression. Our data strongly suggests that the THO complex/Sub2p function is required to coordinate events leading to the acquisition of export competence at a step that follows commitment to 3'-processing.

Impact of a printed decision aid on patients' intention to undergo prostate cancer screening: a multicentre, pragmatic randomised controlled trial in primary care.

BACKGROUND: Despite recommendations against systematic screening for prostate cancer, 70% of patients still request prostate-specific antigen testing. AIM: To assess the impact of a decision aid on patients' intention to undergo prostate cancer screening. DESIGN AND SETTING: Randomised controlled trial with two-arm parallel groups in 86 general practices in urban and rural areas in France. METHOD: Males aged 50-75 years were randomised to receive either the decision aid (intervention group) or usual care (control group). The primary outcome was the proportion of patients' intending to undergo prostate cancer screening, assessed immediately after reading the decision aid. The reasons underlying their choice were elicited and the proportion of patients citing each reason to undergo, or not undergo, prostate cancer screening were compared between the two arms. RESULTS: A total of 1170 patients were randomised (588 in the intervention arm) from November 2012 to February 2013. The proportion of patients who intended to be tested for prostate cancer in the intervention arm (123 patients [20.9%]) was significantly reduced compared with the control arm (57 patients [9.8%]) (difference 11.1%, 95% confidence interval [CI] = 7.0 to 15.2, P<0.0001). In the intervention group, a lower proportion of individuals expressed that cancer screening would protect them from the disease, compared with the control group (P<0.0001), while a greater proportion of individuals stated that prostate cancer screening would not benefit their health (P<0.0001) and may involve procedures with harmful side effects (P = 0.0005). CONCLUSION: The decision aid improved participants' informed decision making and reduced their intent to undergo prostate cancer screening.

Roles for Ctk1 and Spt6 in regulating the different methylation states of histone H3 lysine 36.

Set2 (KMT3)-dependent methylation (me) of histone H3 at lysine 36 (H3K36) promotes deacetylation of transcribed chromatin and represses cryptic promoters within genes. Although Set2 is the only methyltransferase (KMTase) for H3K36 in yeast, it is not known if Set2 is regulated or whether the different methylation states at H3K36 are functionally distinct. Here we show that the N-terminal 261 residues of Set2 (Set2(1-261)), containing the SET KMTase domain, are sufficient for H3K36me2, histone deacetylation, and repression of cryptic promoters at STE11. Set2-catalyzed H3K36me2 does not require either Ctk1-dependent phosphorylation of RNA polymerase II (RNAPII) or the presence of the phospho-C-terminal domain (CTD) interaction (SRI) domain of Set2. This finding is consistent with a known correlation between H3K36me2 and whether a gene is on or off, but not the level of activity of a gene. By contrast, H3K36me3 requires Spt6, proline 38 on histone H3 (H3P38), the CTD of RNAPII, Ctk1, and the C-terminal SRI domain of Set2. We suggest that the C-terminal region of Set2, in conjunction with the phosphorylated CTD of RNAPII, influences the KMTase activity to promote H3K36me3 during transcription elongation.

Transcription termination and nuclear degradation of cryptic unstable transcripts: a role for the nrd1-nab3 pathway in genome surveillance.

Cryptic unstable transcripts (CUTs) are widely distributed in the genome of S. cerevisiae. These RNAs generally derive from nonannotated regions of the genome and are degraded rapidly and efficiently by the nuclear exosome via a pathway that involves degradative polyadenylation by a new poly(A) polymerase borne by the TRAMP complex. What is the share of significant information that is encrypted in CUTs and what distinguishes a CUT from other Pol II transcripts are unclear to date. Here we report the dissection of the molecular mechanism that leads to degradation of a model CUT, NEL025c. We show that the Nrd1p-Nab3p-dependent pathway, involved in transcription termination of sno/snRNAs, is required, albeit not sufficient, for efficient degradation of NEL025c RNAs and at least a subset of other CUTs. Our results suggest an important role for the Nrd1p-Nab3p pathway in the control of gene expression throughout the genome.

Yeast homolog of a cancer-testis antigen defines a new transcription complex.

We have isolated a new yeast gene (PCC1) that codes for a factor homologous to human cancer-testis antigens. We provide evidence that Pcc1p is a new transcription factor and that its mutation affects expression of several genes, some of which are involved in cell cycle progression and polarized growth. Mutation of Pcc1p also affects the expression of GAL genes by impairing the recruitment of the SAGA and Mediator co-activators. We characterize a new complex that contains Pcc1p, a kinase, Bud32p, a putative endopeptidase, Kae1p and two additional proteins encoded by ORFs YJL184w and YMLO36w. Genetic and physical interactions among these proteins strongly suggest that this complex is a functional unit. Chromatin immunoprecipitation experiments and multiple genetic interactions of pcc1 mutants with mutants of the transcription apparatus and chromatin modifying enzymes underscore the direct role of the complex in transcription. Functional complementation experiments indicate that the transcriptional function of this set of genes is conserved throughout evolution.