Abstract

BackgroundRNA polymerase II plays critical roles in transcription in eukaryotic organisms. C-terminal Domain Phosphatase-like 1 (CPL1) regulates the phosphorylation state of the C-terminal domain of RNA polymerase II subunit B1, which is critical in determining RNA polymerase II activity. CPL1 plays an important role in miRNA biogenesis, plant growth and stress responses. Although cpl1 mutant showes delayed-flowering phenotype, the molecular mechanism behind CPL1’s role in floral transition is still unknown.ResultsTo study the role of CPL1 during the floral transition, we first tested phenotypes of cpl1-3 mutant, which harbors a point-mutation. The cpl1-3 mutant contains a G-to-A transition in the second exon, which results in an amino acid substitution from Glu to Lys (E116K). Further analyses found that the mutated amino acid (Glu) was conserved in these species. As a result, we found that the cpl1-3 mutant experienced delayed flowering under both long- and short-day conditions, and CPL1 is involved in the vernalization pathway. Transcriptome analysis identified 109 genes differentially expressed in the cpl1 mutant, with 2 being involved in floral transition. Differential expression of the two flowering-related DEGs was further validated by qRT-PCR.ConclusionsFlowering genetic pathways analysis coupled with transciptomic analysis provides potential genes related to floral transition in the cpl1-3 mutant, and a framework for future studies of the molecular mechanisms behind CPL1’s role in floral transition.

Highlights

  • RNA polymerase II plays critical roles in transcription in eukaryotic organisms

  • Loss of C-terminal Domain Phosphatase-like 1 (CPL1) function delays flowering in Arabidopsis CPL1 regulates flowering time in Arabidopsis [18], but the molecular mechanism underlying CPL1’s role in floral transition is still unknown

  • To confirm the loss of CPL1 function was responsible for the delayed-flowering phenotype of the cpl1-3 mutant, we transformed the cpl1-3 mutant with a construct containing the coding sequence of CPL1 driven by the constitutive cauliflower mosaic virus 35 S promoter

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Summary

Introduction

RNA polymerase II plays critical roles in transcription in eukaryotic organisms. C-terminal Domain Phosphatase-like 1 (CPL1) regulates the phosphorylation state of the C-terminal domain of RNA polymerase II subunit B1, which is critical in determining RNA polymerase II activity. CPL1 plays an important role in miRNA biogenesis, plant growth and stress responses. RNA polymerase II (Pol II) is a multiunit enzyme complex that plays critical roles in transcription in eukaryotic organisms. In Arabidopsis, the CTD phosphatase CPL1 plays an important role in modulating co-transcriptional premRNA processing, thereby affecting growth and stress responses [15]. The phosphorylation state of HYL1, and its activity level, is regulated by CPL1 [13]. RCF3 interacts with CPL1 in the nucleus, and these interactions are essential to regulate the phosphorylation state of HYL1 [19]. The inactivation of RCF3 causes a phosphorylation shift of HYL1 towards the less active version [22]

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