The Nucleic Acid-binding Domain and Translational Repression Activity of a Xenopus Terminal Uridylyl Transferase

Lapointe, Christopher P.; Wickens, Marvin

doi:10.1074/jbc.m113.455451

Cited by 21 publications

(28 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consequently, UTP selectivity of Gld2, which is phylogenetically derived from Tut4/7 (Fig. 5), is conferred by a histidine insertion, rather than an Asn to His mutation as previously suggested (Lunde et al 2012;Yates et al 2012;Lapointe and Wickens 2013).…”

Section: Convergent Evolution Of Tutase Activity By Histidine Insertimentioning

confidence: 78%

“…In Cid1, histidine 336, which is located on a flexible loop near the catalytic site, sterically excludes ATP from the active site. Prior experiments have shown that mutation of this histidine to a smaller amino acid broadens nucleotide specificity in uridylyltransferases to include ATP, concluding that an asparagine to histidine mutation confers ATP specificity (Lunde et al 2012;Yates et al 2012;Lapointe and Wickens 2013). Our multiple sequence alignment ( Fig.…”

Section: Convergent Evolution Of Tutase Activity By Histidine Insertimentioning

confidence: 99%

“…Recent biochemical and structural data on fission yeast Cid1 (Lunde et al 2012;Yates et al 2012) and X. laevis TUT7 (Lapointe and Wickens 2013) suggested that the nucleotide preference of nucleotidyltransferases is determined by a single histidine near the active site (Fig. 6A,B).…”

Section: Convergent Evolution Of Tutase Activity By Histidine Insertimentioning

confidence: 99%

See 2 more Smart Citations

Nucleotide specificity of the human terminal nucleotidyltransferase Gld2 (TUT2)

Chung

Heinemann

2016

RNA

View full text Add to dashboard Cite

The nontemplated addition of single or multiple nucleotides to RNA transcripts is an efficient means to control RNA stability and processing. Cytoplasmic RNA adenylation and the less well-known uridylation are post-transcriptional mechanisms regulating RNA maturation, activity, and degradation. Gld2 is a member of the noncanonical poly(A) polymerases, which include enzymes with varying nucleotide specificity, ranging from strictly ATP to ambiguous to exclusive UTP adding enzymes. Human Gld2 has been associated with transcript stabilizing miRNA monoadenylation and cytoplasmic mRNA polyadenylation. Most recent data have uncovered an unexpected miRNA uridylation activity, which promotes miRNA maturation. These conflicting data raise the question of Gld2 nucleotide specificity. Here, we biochemically characterized human Gld2 and demonstrated that it is a bona fide adenylyltransferase with only weak activity toward other nucleotides. Despite its sequence similarity with uridylyltransferases (TUT4, TUT7), Gld2 displays an 83-fold preference of ATP over UTP. Gld2 is a promiscuous enzyme, with activity toward miRNA, pre-miRNA, and polyadenylated RNA substrates. Apo-Gld2 activity is restricted to adding single nucleotides and processivity likely relies on additional RNA-binding proteins. A phylogeny of the PAP/TUTase superfamily suggests that uridylyltransferases, which are derived from distinct adenylyltransferase ancestors, arose multiple times during evolution via insertion of an active site histidine. A corresponding histidine insertion into the Gld2 active site alters substrate specificity from ATP to UTP.

show abstract

Section: Convergent Evolution Of Tutase Activity By Histidine Insertimentioning

confidence: 78%

Section: Convergent Evolution Of Tutase Activity By Histidine Insertimentioning

confidence: 99%

See 1 more Smart Citation

Nucleotide specificity of the human terminal nucleotidyltransferase Gld2 (TUT2)

Chung

Heinemann

2016

RNA

View full text Add to dashboard Cite

show abstract

“…If storage of mRNAs in P-bodies is regulated by uridylation, deuridylation should be required for reentry of the stored mRNAs into polysomes, suggesting that uridylation is involved in translational inactivation. Indeed, uridylated poly(A) + reporter luciferase RNA is translationally inactive in Xenopus oocytes (Lapointe and Wickens 2013). Therefore, modification of the 3 ′ oligo(U) tail in starfish oocytes may translationally inactivate certain mRNAs prior to hormonal stimulation.…”

Section: Mrna Stabilization and Decay In Oocytesmentioning

confidence: 99%

“…Moreover, uridylation of the 3 ′ end of a polyadenylated luciferase reporter RNA represses translation of the RNA in Xenopus oocytes (Lapointe and Wickens 2013).…”

Section: Introductionmentioning

confidence: 99%

Hormonal stimulation of starfish oocytes induces partial degradation of the 3′ termini of cyclin B mRNAs with oligo(U) tails, followed by poly(A) elongation

Ochi

Chiba

2016

RNA

View full text Add to dashboard Cite

In yeast, plant, and mammalian somatic cells, short poly(A) tails on mRNAs are subject to uridylation, which mediates mRNA decay. Although mRNA uridylation has never been reported in animal oocytes, maternal mRNAs with short poly(A) tails are believed to be translationally repressed. In this study, we found that 96% of cyclin B mRNAs with short poly(A) tails were uridylated in starfish oocytes. Hormonal stimulation induced poly(A) elongation of cyclin B mRNA, and 62% of long adenine repeats did not contain uridine residues. To determine whether uridylated short poly(A) tails destabilize cyclin B mRNA, we developed a method for producing RNAs with the strict 3 ′ terminal sequences of cyclin B, with or without oligo(U) tails. When we injected these synthetic RNAs into starfish oocytes prior to hormonal stimulation, we found that uridylated RNAs were as stable as nonuridylated RNAs. Following hormonal stimulation, the 3 ′ termini of short poly(A) tails of synthesized RNAs containing oligo(U) tails were trimmed, and their poly(A) tails were subsequently elongated. These results indicate that uridylation of short poly(A) tails in cyclin B mRNA of starfish oocytes does not mediate mRNA decay; instead, hormonal stimulation induces partial degradation of uridylated short poly(A) tails in the 3 ′ -5 ′ direction, followed by poly(A) elongation. Oligo(U) tails may be involved in translational inactivation of mRNAs.

show abstract

RNA uridylation: a key posttranscriptional modification shaping the coding and noncoding transcriptome

et al. 2017

View full text Add to dashboard Cite

RNA uridylation is a potent and widespread posttranscriptional regulator of gene expression. RNA uridylation has been detected in a range of eukaryotes including trypanosomes, animals, plants, and fungi, but with the noticeable exception of budding yeast. Virtually all classes of eukaryotic RNAs can be uridylated and uridylation can also tag viral RNAs. The untemplated addition of a few uridines at the 3 0 end of a transcript can have a decisive impact on RNA's fate. In rare instances, uridylation is an intrinsic step in the maturation of noncoding RNAs like for the U6 spliceosomal RNA or mitochondrial guide RNAs in trypanosomes. Uridylation can also switch specific miRNA precursors from a degradative to a processing mode. This switch depends on the number of uridines added which is regulated by the cellular context. Yet, the typical consequence of uridylation on mature noncoding RNAs or their precursors is to accelerate decay. Importantly, mRNAs are also tagged by uridylation. In fact, the advent of novel high throughput sequencing protocols has recently revealed the pervasiveness of mRNA uridylation, from plants to humans. As for noncoding RNAs, the main function to date for mRNA uridylation is to promote degradation. Yet, additional roles begin to be ascribed to U-tailing such as the control of mRNA deadenylation, translation control and possibly storage. All these new findings illustrate that we are just beginning to appreciate the diversity of roles played by RNA uridylation and its full temporal and spatial implication in regulating gene expression.

show abstract

The Nucleic Acid-binding Domain and Translational Repression Activity of a Xenopus Terminal Uridylyl Transferase

Cited by 21 publications

References 69 publications

Nucleotide specificity of the human terminal nucleotidyltransferase Gld2 (TUT2)

Nucleotide specificity of the human terminal nucleotidyltransferase Gld2 (TUT2)

Hormonal stimulation of starfish oocytes induces partial degradation of the 3′ termini of cyclin B mRNAs with oligo(U) tails, followed by poly(A) elongation

RNA uridylation: a key posttranscriptional modification shaping the coding and noncoding transcriptome

Contact Info

Product

Resources

About