Antisense-mediated FLC transcriptional repression requires the P-TEFb transcription elongation factor

Proc Natl Acad Sci U S A. 2014 May 20;111(20):7468-73. doi: 10.1073/pnas.1406635111. Epub 2014 May 5.

Abstract

The functional significance of noncoding transcripts is currently a major question in biology. We have been studying the function of a set of antisense transcripts called COOLAIR that encompass the whole transcription unit of the Arabidopsis floral repressor FLOWERING LOCUS C (FLC). Alternative polyadenylation of COOLAIR transcripts correlates with different FLC sense expression states. Suppressor mutagenesis aimed at understanding the importance of this sense-antisense transcriptional circuitry has identified a role for Arabidopsis cyclin-dependent kinase C (CDKC;2) in FLC repression. CDKC;2 functions in an Arabidopsis positive transcription elongation factor b (P-TEFb) complex and influences global RNA polymerase II (Pol II) Ser(2) phosphorylation levels. CDKC;2 activity directly promotes COOLAIR transcription but does not affect an FLC transgene missing the COOLAIR promoter. In the endogenous gene context, however, the reduction of COOLAIR transcription by cdkc;2 disrupts a COOLAIR-mediated repression mechanism that increases FLC expression. This disruption then feeds back to indirectly increase COOLAIR expression. This tight interconnection between sense and antisense transcription, together with differential promoter sensitivity to P-TEFb, is central to quantitative regulation of this important floral repressor gene.

Keywords: autonomous pathway; chromatin silencing; lncRNA; transcriptional regulation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Chromatin / metabolism
  • Cyclin-Dependent Kinases / metabolism
  • Gene Expression Regulation, Plant*
  • Gene Silencing
  • Genetic Complementation Test
  • MADS Domain Proteins / genetics*
  • Mutation
  • Oligonucleotides, Antisense / genetics*
  • Phosphorylation
  • Plants, Genetically Modified
  • Positive Transcriptional Elongation Factor B / metabolism*
  • Promoter Regions, Genetic
  • Repressor Proteins / genetics
  • Transgenes

Substances

  • Arabidopsis Proteins
  • Chromatin
  • FLF protein, Arabidopsis
  • MADS Domain Proteins
  • Oligonucleotides, Antisense
  • Repressor Proteins
  • Positive Transcriptional Elongation Factor B
  • CDKC;2 protein, Arabidopsis
  • Cyclin-Dependent Kinases