Related Experiment Video
Updated: Dec 30, 2025
10:52
Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions
Published on: September 28, 2017
8.5K
Characterization of METTL16 as a cytoplasmic RNA binding protein
Daniel J Nance1, Emily R Satterwhite1, Brinda Bhaskar1
1Department of Biochemistry and Molecular Biology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, United States of America.
Plos One
|January 16, 2020
Summary
METTL16, an RNA methyltransferase, is found in the cytoplasm, not just the nucleus. This cytoplasmic localization may influence its RNA targets and gene expression regulation.
Area of Science:
- Molecular Biology
- Epigenetics
- RNA Biology
Background:
- N6-methyladenosine (m6A) modification regulates mRNA stability, splicing, and translation.
- The METTL3-METTL14-WTAP complex is a known m6A methyltransferase.
- METTL16 is identified as an RNA m6A methyltransferase, but its biological role is unclear.
Purpose of the Study:
- To identify and verify RNA targets of METTL16.
- To investigate the cellular localization of METTL16.
- To explore the functional role of METTL16 in gene expression.
Main Methods:
- Immunoprecipitation of endogenous and exogenous FLAG-tagged METTL16.
- Biochemical fractionation to determine protein localization.
- siRNA knockdown of METTL16 to assess gene expression changes.
Main Results:
- METTL16 targets were identified and verified.
- Exogenous METTL16 showed different RNA target affinities compared to endogenous METTL16.
- Biochemical fractionation revealed METTL16 predominantly resides in the cytoplasm.
- METTL16 knockdown led to changes in mRNA target expression.
Conclusions:
- METTL16 functions as a cytoplasmic RNA methyltransferase, in addition to its previously reported nuclear role.
- Cellular localization may influence METTL16's RNA binding preferences.
- METTL16 plays a role in regulating gene expression.
Related Concept Videos
01:29Translational Regulation
469
Translational regulation in prokaryotes ensures efficient protein synthesis by controlling ribosome access to mRNA. This regulation is mediated by secondary RNA structures, including translational riboswitches, RNA thermometers, and small RNAs (sRNAs), which respond to intracellular and environmental signals to modulate gene expression.Translational RiboswitchesRiboswitches in the leader region of mRNAs can regulate translation by altering the accessibility of the Shine-Dalgarno (SD) sequence,...
469
01:56Riboswitches
9.4K
Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...
9.4K
01:53RNA Stability
35.5K
Intact DNA strands can be found in fossils, while scientists sometimes struggle to keep RNA intact under laboratory conditions. The structural variations between RNA and DNA underlie the differences in their stability and longevity. Because DNA is double-stranded, it is inherently more stable. The single-stranded structure of RNA is less stable but also more flexible and can form weak internal bonds. Additionally, most RNAs in the cell are relatively short, while DNA can be up to 250 million...
35.5K
01:23Types of RNA
72.3K
Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...
72.3K
02:24Ribosome Profiling
4.0K
Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...
4.0K
02:28Leaky Scanning
5.6K
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.6K