ABSTRACT: The human CCHC-type Zinc Finger nucleic acid binding protein binds G-rich elements in target mRNA coding sequences initiation promotes translation (Ribosome profiling).
Project description:The human cellular nucleic acid binding protien binds G-rich elements close to translation initiation sires and promotes translation.
Project description:The human cellular nucleic acid binding protien binds G-rich elements close to translation initiation sires and promotes translation. [PAR-CLIP]
Project description:The human cellular nucleic acid binding protien binds G-rich elements close to translation initiation sires and promotes translation. [RNA-Seq]
Project description:Protein translation is tightly regulated at the initiation phase, which entails exceedingly complex interactions among multiple EIF initiation factors to bring the ribosome to the mRNA and scan the 5’ leader sequence for the start codon. EIF-3 contains multiple subunits with essential and emerging regulatory activities throughout the translation initiation pathway. However, it remains poorly understood how individual components of the EIF3 complex contribute to the selective regulation of translation in multicellular organisms. Here, through studies of a missense mutation in C. elegans EIF-3.G, an RNA-binding subunit of EIF3, we have dissected mechanisms of how regulation of protein translation initiation modulates neuronal excitation. EIF-3.G(C130Y) alters a conserved Zinc Finger and behaves as a gain-of-function. We show that while EIF-3.G(C130Y) permits essential aspects of translation initiation, this variant functionally involves its RNA binding activity in regulating protein synthesis. We mapped EIF-3.G binding sites in the C. elegans cholinergic motor neuron transcriptome and identified a significant representation of mRNAs containing long and GC-rich 5' UTRs and in mRNAs exhibiting activity-dependent transcript level changes. Our results suggest that modulation of the scanning process of translation initiation by EIF3 contributes homeostatic regulation to synaptic activity changes.
Project description:CNBP is a eukaryote-conserved nucleic-acid binding protein required in mammals for embryonic development. It contains seven CCHC-type zinc-finger domains and was suggested to act as a nucleic acid chaperone, as well as a transcription factor. Here, we identify all CNBP isoforms as cytoplasmic messenger RNA (mRNA)-binding proteins. Using Photoactivatable Ribonucleoside Enhanced Cross-linking and Immunoprecipitation, we mapped its binding sites on RNA at nucleotide-level resolution on a genome-wide scale and find that CNBP interacted with 3961 mRNAs in human cell lines, preferentially at a G-rich motif close to the AUG start codon on mature mRNAs. Loss- and gain-of-function analyses coupled with system-wide RNA and protein quantification revealed that CNBP did not affect RNA abundance, but rather promoted translation of its targets. This is consistent with an RNA chaperone function of CNBP helping to resolve secondary structures, thus promoting translation. CNBP PAR-CLIP
Project description:CNBP is a eukaryote-conserved nucleic-acid binding protein required in mammals for embryonic development. It contains seven CCHC-type zinc-finger domains and was suggested to act as a nucleic acid chaperone, as well as a transcription factor. Here, we identify all CNBP isoforms as cytoplasmic messenger RNA (mRNA)-binding proteins. Using Photoactivatable Ribonucleoside Enhanced Cross-linking and Immunoprecipitation, we mapped its binding sites on RNA at nucleotide-level resolution on a genome-wide scale and find that CNBP interacted with 3961 mRNAs in human cell lines, preferentially at a G-rich motif close to the AUG start codon on mature mRNAs. Loss- and gain-of-function analyses coupled with system-wide RNA and protein quantification revealed that CNBP did not affect RNA abundance, but rather promoted translation of its targets. This is consistent with an RNA chaperone function of CNBP helping to resolve secondary structures, thus promoting translation. CNBP protein knockdown and RNA-seq
Project description:CNBP is a eukaryote-conserved nucleic-acid binding protein required in mammals for embryonic development. It contains seven CCHC-type zinc-finger domains and was suggested to act as a nucleic acid chaperone, as well as a transcription factor. Here, we identify all CNBP isoforms as cytoplasmic messenger RNA (mRNA)-binding proteins. Using Photoactivatable Ribonucleoside Enhanced Cross-linking and Immunoprecipitation, we mapped its binding sites on RNA at nucleotide-level resolution on a genome-wide scale and find that CNBP interacted with 3961 mRNAs in human cell lines, preferentially at a G-rich motif close to the AUG start codon on mature mRNAs. Loss- and gain-of-function analyses coupled with system-wide RNA and protein quantification revealed that CNBP did not affect RNA abundance, but rather promoted translation of its targets. This is consistent with an RNA chaperone function of CNBP helping to resolve secondary structures, thus promoting translation.
Project description:CNBP is a eukaryote-conserved nucleic-acid binding protein required in mammals for embryonic development. It contains seven CCHC-type zinc-finger domains and was suggested to act as a nucleic acid chaperone, as well as a transcription factor. Here, we identify all CNBP isoforms as cytoplasmic messenger RNA (mRNA)-binding proteins. Using Photoactivatable Ribonucleoside Enhanced Cross-linking and Immunoprecipitation, we mapped its binding sites on RNA at nucleotide-level resolution on a genome-wide scale and find that CNBP interacted with 3961 mRNAs in human cell lines, preferentially at a G-rich motif close to the AUG start codon on mature mRNAs. Loss- and gain-of-function analyses coupled with system-wide RNA and protein quantification revealed that CNBP did not affect RNA abundance, but rather promoted translation of its targets. This is consistent with an RNA chaperone function of CNBP helping to resolve secondary structures, thus promoting translation.
