A New Marker for Mosaic Analysis in Caenorhabditis elegans Indicates a Fusion Between hyp6 and hyp7, Two Major Components of the Hypodermis

Yochem, John; Gu, Trent; Han, Min

doi:10.1093/genetics/149.3.1323

Cited by 193 publications

(5 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While it has been suggested that individual importins act as receptors for several different types of protein cargo, 47 we reasoned that AKIR-1 is likely involved in a more limited role. We therefore used a nuclear localized GFP transgenic reporter strain ( sur-5p::NLS-GFP ) 33 , 48 to investigate whether AKIR-1 affects nuclear import in general. As expected, RNAi against ima-3 markedly reduced the GFP signal in intestinal nuclei, but akir-1 RNAi had no effect on the nuclear localization of the GFP reporter suggesting a more specific role for akir-1 in nuclear import than for importins ( Figure 7 E).…”

Section: Resultsmentioning

confidence: 99%

AKIR-1 regulates proteasome subcellular function in Caenorhabditis elegans

Pispa,

Mikkonen,

Arpalahti

et al. 2023

iScience

View full text Add to dashboard Cite

Section: Resultsmentioning

confidence: 99%

AKIR-1 regulates proteasome subcellular function in Caenorhabditis elegans

Pispa,

Mikkonen,

Arpalahti

et al. 2023

iScience

View full text Add to dashboard Cite

“…Whilst it has been suggested that individual importins act as receptors for several different types of protein cargo (Kimura et al, 2017), we reasoned that AKIR-1 is likely involved in a more limited role. We therefore used a nuclear localized GFP transgenic reporter strain ( sur-5p::NLS-GFP ) (Yochem et al, 1998; Mikkonen et al, 2017) to investigate whether AKIR-1 affects nuclear import in general. As expected, RNAi against ima-3 markedly reduced the GFP signal in intestinal nuclei, but akir-1 RNAi had no effect on the nuclear localization of the GFP reporter suggesting a more specific role for akir-1 in nuclear import than for importins (Figure 9E).…”

Section: Resultsmentioning

confidence: 99%

AKIR-1 Regulates Proteasome Localization and Function inCaenorhabditis elegans

Pispa

Mikkonen

Arpalahti

et al. 2022

Preprint

View full text Add to dashboard Cite

Regulated protein clearance is vital for cells to maintain protein homeostasis and the conditions essential for survival. The primary machinery for intracellular protein degradation is the ubiquitin –proteasome system (UPS), by which ubiquitin−tagged proteins are degraded by the proteasome. Proteasomes are present both in the cytoplasm and the nucleus, but the mechanisms coordinating proteasome activity and its subcellular localization in a multicellular organism are still unclear. Here, we identified the nuclear protein−encoding gene akir−1 as a proteasome regulator in a genome−wide Caenorhabditis elegans (C. elegans) RNAi screen. We show that the depletion of akir−1 causes accumulation of endogenous polyubiquitinated proteins in the nuclei of intestinal cells, concomitant with slower in vivo proteasomal degradation in this subcellular compartment. Remarkably, the loss of akir−1 does not induce an accumulation of polyubiquitinated proteins in oocyte nuclei, though akir−1 is essential for the nuclear localization of proteasomes in both cell types. We further show that the importin family member ima−3 genetically interacts with akir−1, and affects subcellular distribution of polyubiquitinated proteins in intestinal cells. We show for the first time that conserved AKIR−1 is important for the nuclear transport of proteasomes in a multicellular organism, suggesting a role for AKIR−1 in the maintenance of proteostasis.

show abstract

“…Incorporation at the UAGA codon thus results in production of a GFP::mCherry fusion protein that localizes to the cell nucleus due to the C-terminal NLS. Expression of the protein coding genes was driven by the ubiquitous sur-5p promoter ( 34 ). To construct strains, we used a genetic background containing a deletion in the smg-6 gene, knocking out the nonsense mediated decay machinery ( 35 ).…”

Section: Resultsmentioning

confidence: 99%

Using a quadruplet codon to expand the genetic code of an animal

Davis

Baxter

et al. 2021

Nucleic Acids Research

View full text Add to dashboard Cite

Genetic code expansion in multicellular organisms is currently limited to the use of repurposed amber stop codons. Here, we introduce a system for the use of quadruplet codons to direct incorporation of non-canonical amino acids in vivo in an animal, the nematode worm Caenorhabditis elegans. We develop hybrid pyrrolysyl tRNA variants to incorporate non-canonical amino acids in response to the quadruplet codon UAGA. We demonstrate the efficiency of the quadruplet decoding system by incorporating photocaged amino acids into two proteins widely used as genetic tools. We use photocaged lysine to express photocaged Cre recombinase for the optical control of gene expression and photocaged cysteine to express photo-activatable caspase for light inducible cell ablation. Our approach will facilitate the routine adoption of quadruplet decoding for genetic code expansion in eukaryotic cells and multicellular organisms.

show abstract

A New Marker for Mosaic Analysis in Caenorhabditis elegans Indicates a Fusion Between hyp6 and hyp7, Two Major Components of the Hypodermis

Cited by 193 publications

References 23 publications

AKIR-1 regulates proteasome subcellular function in Caenorhabditis elegans

AKIR-1 regulates proteasome subcellular function in Caenorhabditis elegans

AKIR-1 Regulates Proteasome Localization and Function inCaenorhabditis elegans

Using a quadruplet codon to expand the genetic code of an animal

Contact Info

Product

Resources

About