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STATEMENT 1 Proper localization and functions of macromolecules during cell division are crucial to 2 ensure survival and proliferation of daughter cells. 3 4 ABSTRACT 1 Staufen1 (STAU1) is an RNA-binding protein involved in the posttranscriptional 2 regulation of mRNAs. We report that a large fraction of STAU1 localizes to the mitotic 3 spindle in the colorectal cancer HCT116 and in the non-transformed hTERT-RPE1 cells. 4Spindle-associated STAU1 partly co-localizes with ribosomes and active sites of 5 translation. We mapped the molecular determinant required for STAU1/spindle association 6 within the first 88 N-terminal amino acids, a domain that is not required for RNA binding. 7Interestingly, transcriptomic analysis of purified mitotic spindles reveals that 1054 mRNAs 8 as well as the precursor ribosomal RNA and lncRNAs and snoRNAs involved in 9 ribonucleoprotein assembly and processing are enriched on spindles compared to cell 10 extracts. STAU1 knockout causes the displacement of the pre-rRNA and of 154 mRNAs 11 coding for proteins involved in actin cytoskeleton organization and cell growth, 12 highlighting a role for STAU1 in mRNA trafficking to spindle. These data demonstrate 13 that STAU1 controls the localization of sub-populations of RNAs during mitosis and 14 suggests a novel role of STAU1 in pre-rRNA maintenance during mitosis, ribogenesis 15 and/or nucleoli reassembly. 16 17 18
STATEMENT 1 Proper localization and functions of macromolecules during cell division are crucial to 2 ensure survival and proliferation of daughter cells. 3 4 ABSTRACT 1 Staufen1 (STAU1) is an RNA-binding protein involved in the posttranscriptional 2 regulation of mRNAs. We report that a large fraction of STAU1 localizes to the mitotic 3 spindle in the colorectal cancer HCT116 and in the non-transformed hTERT-RPE1 cells. 4Spindle-associated STAU1 partly co-localizes with ribosomes and active sites of 5 translation. We mapped the molecular determinant required for STAU1/spindle association 6 within the first 88 N-terminal amino acids, a domain that is not required for RNA binding. 7Interestingly, transcriptomic analysis of purified mitotic spindles reveals that 1054 mRNAs 8 as well as the precursor ribosomal RNA and lncRNAs and snoRNAs involved in 9 ribonucleoprotein assembly and processing are enriched on spindles compared to cell 10 extracts. STAU1 knockout causes the displacement of the pre-rRNA and of 154 mRNAs 11 coding for proteins involved in actin cytoskeleton organization and cell growth, 12 highlighting a role for STAU1 in mRNA trafficking to spindle. These data demonstrate 13 that STAU1 controls the localization of sub-populations of RNAs during mitosis and 14 suggests a novel role of STAU1 in pre-rRNA maintenance during mitosis, ribogenesis 15 and/or nucleoli reassembly. 16 17 18
The last stage of cell division involves two daughter cells remaining interconnected by a cytokinetic bridge that is cleaved in a process called abscission. During pre-abscission, we identified that the centrosome moves in a Rab11-dependent manner towards the cytokinetic bridge in human cells grown in culture and in an in vivo vertebrate model, Danio rerio (zebrafish). Rab11-endosomes are dynamically organized in a Rab11-GTP dependent manner at the centrosome during pre-abscission and this organization is required for the centrosome protein, pericentrin, to be enriched at the centrosome. Using zebrafish embryos, we found that reduction in pericentrin expression or optogenetically disrupting Rab11-endosome function inhibited centrosome movement towards the cytokinetic bridge and abscission resulting in daughter cells prone to being binucleated and/or having supernumerary centrosomes. These studies suggest that Rab11-endosomes contribute to centrosome function during pre-abscission by regulating pericentrin organization resulting in appropriate centrosome movement towards the cytokinetic bridge and subsequent abscission.
Staufen1 (STAU1) is an RNA-binding protein involved in the post-transcriptional regulation of mRNAs. We report that a large fraction of STAU1 localizes to the mitotic spindle in colorectal cancer HCT116 cells and in non-transformed hTERT-RPE1 cells. Spindle-associated STAU1 partly co-localizes with ribosomes and active sites of translation. We mapped the molecular determinant required for STAU1–spindle association within the first 88 N-terminal amino acids, a domain that is not required for RNA binding. Interestingly, transcriptomic analysis of purified mitotic spindles revealed that 1054 mRNAs and the precursor ribosomal RNA (pre-rRNA), as well as the long non-coding RNAs and small nucleolar RNAs involved in ribonucleoprotein assembly and processing, are enriched on spindles compared with cell extracts. STAU1 knockout causes displacement of the pre-rRNA and of 154 mRNAs coding for proteins involved in actin cytoskeleton organization and cell growth, highlighting a role for STAU1 in mRNA trafficking to spindle. These data demonstrate that STAU1 controls the localization of subpopulations of RNAs during mitosis and suggests a novel role of STAU1 in pre-rRNA maintenance during mitosis, ribogenesis and/or nucleoli reassembly.
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