Abstract

Key messageHere we demonstrate that the APUM9 RNA-binding protein and its co-factors play a role in mRNA destabilization and how this activity might regulate early plant development.APUM9 is a conserved PUF RNA-binding protein (RBP) under complex transcriptional control mediated by a transposable element (TE) that restricts its expression in Arabidopsis. Currently, little is known about the functional and mechanistic details of the plant PUF regulatory system and the biological relevance of the TE-mediated repression of APUM9 in plant development and stress responses. By combining a range of transient assays, we show here, that APUM9 binding to target transcripts can trigger their rapid decay via its conserved C-terminal RNA-binding domain. APUM9 directly interacts with DCP2, the catalytic subunit of the decapping complex and DCP2 overexpression induces rapid decay of APUM9 targeted mRNAs. We show that APUM9 negatively regulates the expression of ABA signaling genes during seed imbibition, and thereby might contribute to the switch from dormant stage to seed germination. By contrast, strong TE-mediated repression of APUM9 is important for normal plant growth in the later developmental stages. Finally, APUM9 overexpression plants show slightly enhanced heat tolerance suggesting that TE-mediated control of APUM9, might have a role not only in embryonic development, but also in plant adaptation to heat stress conditions.

Highlights

  • The Arabidopsis PUMILIO9 (APUM9) protein is a member of the highly conserved family of PUF RNA-binding proteins found in all Accession number The RNA seq data generated in this study are available in the GEO database under GSE104860

  • If APUM9 tethering leads to mRNA destabilization, co-infiltration of λN-APUM9 without λN fusion (A9) but not λN or A9 should result in a decrease of the transcript and protein levels of GFP5BB, while translational repression by APUM9 should only influence GFP5BB protein accumulation

  • In order to analyze whether the non-conserved N-terminal or the highly conserved C-terminal part of APUM9 is required for target repression, the λN-A9Nt and λN-A9Ct constructs, expressing only the N, or C-terminal APUM9 domains were co-infiltrated with GFP5BB (Fig. 1b)

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Summary

Introduction

The Arabidopsis PUMILIO9 (APUM9) protein is a member of the highly conserved family of PUF RNA-binding proteins (referred as PUMILIO proteins) found in all Accession number The RNA seq data generated in this study are available in the GEO database under GSE104860. Conserved interactions between PUF proteins and POP2 deadenylase subunit are well described from yeast to mammals the precise decay mechanism(s) remains to be unraveled (Goldstrohm et al 2006). Protein partners can tune PUF activity in a tissue or developmentspecific manner leading to altered RNA-binding affinity or change of affinity to target mRNAs (Wickens et al 2002; Weidmann et al 2016). A conserved long noncoding RNA (lncRNA), NORAD, can serve as a molecular decoy and control the activity of PUM1–2 PUF proteins to preserve genome stability (Lee et al 2016; Tichon et al 2016)

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