Efficient globin production during terminal erythropoiesis depends on the synergistic action of TENT5C poly(A) polymerase and LARP4/5

<p><span><span>Red blood cell development is a unique process </span><span>where</span> <span>reduced</span> <span>transcriptome and proteome complexity</span> <span>facilitate</span><span>s</span><span> vast</span> <span>hemoglobin</span><span> production</span><span>. Here, we describe the cooperative role of cytoplasmic poly(A) polymerase </span><span>TENT5C </span><span>and the poly(A) tail-protecting </span><span>LARP4</span><span>/5 RNA-binding proteins in ensuring proper </span><span>hemoglobin production</span><span>. </span><span>TENT5C </span><span>catalytic mutant </span><span>knock-in </span><span>mice</span><span> display microcytic hypochromic anemia </span><span>resembling</span><span> constitutive knockout</span><span>. </span><span>TENT5C </span><span>counteracts gradual globin mRNA </span><span>deadenylation</span> <span>during</span><span> erythropoiesis. </span><span>In the late stages, TENT5C</span><span> dysfunction leads to globin poly(A) tail shortening and a </span><span>drastic reduction of mRNA</span> <span>level</span><span>s</span> <span>in reticulocytes. </span><span>P</span><span>roteomic experiments revealed </span><span>transient </span><span>but</span><span> specific</span><span> association of TENT5C with </span><span>LARP4</span><span>/5</span><span>. </span><span>Indeed</span><span>,</span><span> LARP</span><span>4/</span><span>5</span> <span>depletion </span><span>leads to downregulation and poly(A) tail shortening of globin mRNAs</span><span>.</span><span> Furthermore</span><span>,</span> <span>lack of TENT5C catalytic activity </span><span>is accompanied by </span><span>compensatory </span><span>upregulation </span><span>of </span><span>LARP4</span><span>/</span><span>5. </span><span>F</span><span>i</span><span>nally</span><span>, the</span><span> importance of</span><span> precise regulation of globin poly(A) tails </span><span>by </span><span>deadenylation</span><span> and re-adenylation </span><span>is highlighted by the destabilization of TENT5C </span><span>by</span><span> CCR4-</span><span>NOT </span><span>deadenylase</span><span> complex-associated </span><span>E3 </span><span>ubiquitin ligase </span><span>C</span><span>NOT4.</span></span><span>&nbsp;<br><br>This repository contains supplementary data accompanying related publication. The uploaded files contain following data:<br><br></span></p> <div> <p><span><strong><span>Supplementary </span><span>Data </span></strong><span><strong>1</strong> </span></span><span><span>DRS of E14.5 fetal livers, </span><span>poly(A) </span><span>profiling </span><span>stat</span><span>istics.</span></span><span>&nbsp;</span></p> </div> <div> <p><span><strong><span>Supplementary </span><span>Data </span><span>2</span></strong> </span><span><span>(A) All </span><span>identified</span><span> proteins </span><span>in the</span> <span>TurboID</span><span> assay </span><span>(B) Overlap between 3 replicates (C) Statistically significant GO terms for overlapping proteins with average specificity &gt; 0.</span></span><span>&nbsp;</span></p> </div> <div> <p><span><strong><span>Supplementary </span><span>Data</span> <span>3</span></strong> </span><span><span>(A) Overview of used input and method details used in </span><span>degronopedia</span><span> analysis (B) Identified degron motifs (C) Degron conservation (D) Evolutionary conservation of TENT5C aa sequence scored by </span><span>ConSurf</span><span> (The table shows the residue variety in % for each position in the query sequence, each column shows the % for that amino acid found in position in the MSA)</span><span>.</span></span><span>&nbsp;</span></p> </div> <div> <p><span><strong><span>Supplementary </span><span>Data </span></strong><span><strong>4</strong> </span></span><span><span>Key resources table</span><span>.</span></span><span>&nbsp;</span></p> </div> <div> <p><span><span><strong>Supplementary Data 5</strong> </span></span><span><span>Oligonucleotides </span><span>used</span><span> in the study</span><span>.</span></span><span>&nbsp;</span></p> </div> <div> <p><strong><span><span>Supplementary Resource</span><span> 1</span> </span></strong><span><span>TurboID</span><span> assay</span></span> <span><span>MS/MS raw data</span><span>, each file in the</span><span> archive</span><span> corresponds to a </span><span>biological replicate of either WT (control) or </span></span><span><span>Tent5c-EGFP</span><span>.</span></span><span>&nbsp;</span></p> </div> <p><span>&nbsp;</span></p>