Frenk Stephen, Oxley David, Houseley Jonathan
Epigenetics Programme, The Babraham Institute, Cambridge, United Kingdom.
Proteomics Group, The Babraham Institute, Cambridge, United Kingdom.
PLoS One. 2014 Sep 11;9(9):e107648. doi: 10.1371/journal.pone.0107648. eCollection 2014.
Nuclear RNA degradation pathways are highly conserved across eukaryotes and play important roles in RNA quality control. Key substrates for exosomal degradation include aberrant functional RNAs and cryptic unstable transcripts (CUTs). It has recently been reported that the nuclear exosome is inactivated during meiosis in budding yeast through degradation of the subunit Rrp6, leading to the stabilisation of a subset of meiotic unannotated transcripts (MUTs) of unknown function. We have analysed the activity of the nuclear exosome during meiosis by deletion of TRF4, which encodes a key component of the exosome targeting complex TRAMP. We find that TRAMP mutants produce high levels of CUTs during meiosis that are undetectable in wild-type cells, showing that the nuclear exosome remains functional for CUT degradation, and we further report that the meiotic exosome complex contains Rrp6. Indeed Rrp6 over-expression is insufficient to suppress MUT transcripts, showing that the reduced amount of Rrp6 in meiotic cells does not directly cause MUT accumulation. Lack of TRAMP activity stabilises ∼ 1600 CUTs in meiotic cells, which occupy 40% of the binding capacity of the nuclear cap binding complex (CBC). CBC mutants display defects in the formation of meiotic double strand breaks (DSBs), and we see similar defects in TRAMP mutants, suggesting that a key function of the nuclear exosome is to prevent saturation of the CBC complex by CUTs. Together, our results show that the nuclear exosome remains active in meiosis and has an important role in facilitating meiotic recombination.
核RNA降解途径在真核生物中高度保守,在RNA质量控制中发挥重要作用。外泌体降解的关键底物包括异常功能RNA和隐蔽不稳定转录本(CUTs)。最近有报道称,在芽殖酵母减数分裂过程中,核外泌体通过亚基Rrp6的降解而失活,导致一部分功能未知的减数分裂未注释转录本(MUTs)稳定。我们通过缺失TRF4(其编码外泌体靶向复合物TRAMP的关键组分)分析了减数分裂过程中核外泌体的活性。我们发现,TRAMP突变体在减数分裂过程中产生大量野生型细胞中无法检测到的CUTs,这表明核外泌体对CUT降解仍有功能,并且我们进一步报道减数分裂外泌体复合物含有Rrp6。实际上,Rrp6过表达不足以抑制MUT转录本,这表明减数分裂细胞中Rrp6量的减少不会直接导致MUT积累。缺乏TRAMP活性会使减数分裂细胞中约1600种CUTs稳定,这些CUTs占据了核帽结合复合物(CBC)结合能力的40%。CBC突变体在减数分裂双链断裂(DSBs)形成方面表现出缺陷,我们在TRAMP突变体中也观察到类似缺陷,这表明核外泌体的一个关键功能是防止CUTs使CBC复合物饱和。总之,我们的结果表明核外泌体在减数分裂中保持活性,并且在促进减数分裂重组中起重要作用。