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Novel origin of lamin-derived cytoplasmic intermediate filaments in tardigrades.

Hering L, Bouameur JE, Reichelt J, Magin TM, Mayer G - Elife (2016)

Bottom Line: Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods.Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades.These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Institute of Biology, University of Kassel, Kassel, Germany.

ABSTRACT
Intermediate filament (IF) proteins, including nuclear lamins and cytoplasmic IF proteins, are essential cytoskeletal components of bilaterian cells. Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods. Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades. Transcriptomic and genomic data revealed three IF protein genes in the tardigrade Hypsibius dujardini, one of which (cytotardin) occurs exclusively in the cytoplasm of epidermal and foregut epithelia, where it forms belt-like filaments around each epithelial cell. These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians.

No MeSH data available.


Plasmids, primers, and restriction enzymes used for the cloning of tardigrade lamin and cytotardin genes.DOI:http://dx.doi.org/10.7554/eLife.11117.017
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fig5s2: Plasmids, primers, and restriction enzymes used for the cloning of tardigrade lamin and cytotardin genes.DOI:http://dx.doi.org/10.7554/eLife.11117.017

Mentions: Total RNA from several hundred specimens of H. dujardini was extracted and purified using TRIzol Reagent (Life Technologies, Carlsbad, CA) and RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany) according to the manufacturers’ protocols. First strand cDNA synthesis was performed using random hexamer primer and SuperScript II Reverse Transcriptase (Life Technologies) and afterwards used as template for amplification of the whole coding sequence (CDS) of lamin-1, lamin-2 and cytotardin using gene specific primers. The cytotardin specific primers contained restriction sites that were required for subsequent cloning into bacterial or mammalian expression vectors (see Figure 5—figure supplement 2). The amplicons of lamin-1 and lamin-2 were cloned into the pGEM-T Vector System (Promega, Madison, WI) to generate the plasmids pGEM-T-lamin-1 and pGEM-T-lamin-2. Cytotardin CDS was cloned into expression vectors pET15b (Novagen Merck Millipore, Darmstadt, Germany), pcDNA3.1/Zeo (+) (Life Technologies) and pEGFP-C3 (Clontech Laboratories, Mountain View, CA) to generate the following plasmids: pET15b-cytotardin, pcDNA3-cytotardin, pcDNA3-HA-cytotardin and pEGFP-cytotardin. All PCR-amplified constructs were verified by Sanger sequencing and have been deposited in GenBank (http://www.ncbi.nlm.nih.gov/genbank) under accession numbers KU295460–KU295467.


Novel origin of lamin-derived cytoplasmic intermediate filaments in tardigrades.

Hering L, Bouameur JE, Reichelt J, Magin TM, Mayer G - Elife (2016)

Plasmids, primers, and restriction enzymes used for the cloning of tardigrade lamin and cytotardin genes.DOI:http://dx.doi.org/10.7554/eLife.11117.017
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4829535&req=5

fig5s2: Plasmids, primers, and restriction enzymes used for the cloning of tardigrade lamin and cytotardin genes.DOI:http://dx.doi.org/10.7554/eLife.11117.017
Mentions: Total RNA from several hundred specimens of H. dujardini was extracted and purified using TRIzol Reagent (Life Technologies, Carlsbad, CA) and RNeasy MinElute Cleanup Kit (Qiagen, Hilden, Germany) according to the manufacturers’ protocols. First strand cDNA synthesis was performed using random hexamer primer and SuperScript II Reverse Transcriptase (Life Technologies) and afterwards used as template for amplification of the whole coding sequence (CDS) of lamin-1, lamin-2 and cytotardin using gene specific primers. The cytotardin specific primers contained restriction sites that were required for subsequent cloning into bacterial or mammalian expression vectors (see Figure 5—figure supplement 2). The amplicons of lamin-1 and lamin-2 were cloned into the pGEM-T Vector System (Promega, Madison, WI) to generate the plasmids pGEM-T-lamin-1 and pGEM-T-lamin-2. Cytotardin CDS was cloned into expression vectors pET15b (Novagen Merck Millipore, Darmstadt, Germany), pcDNA3.1/Zeo (+) (Life Technologies) and pEGFP-C3 (Clontech Laboratories, Mountain View, CA) to generate the following plasmids: pET15b-cytotardin, pcDNA3-cytotardin, pcDNA3-HA-cytotardin and pEGFP-cytotardin. All PCR-amplified constructs were verified by Sanger sequencing and have been deposited in GenBank (http://www.ncbi.nlm.nih.gov/genbank) under accession numbers KU295460–KU295467.

Bottom Line: Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods.Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades.These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians.

View Article: PubMed Central - PubMed

Affiliation: Department of Zoology, Institute of Biology, University of Kassel, Kassel, Germany.

ABSTRACT
Intermediate filament (IF) proteins, including nuclear lamins and cytoplasmic IF proteins, are essential cytoskeletal components of bilaterian cells. Despite their important role in protecting tissues against mechanical force, no cytoplasmic IF proteins have been convincingly identified in arthropods. Here we show that the ancestral cytoplasmic IF protein gene was lost in the entire panarthropod (onychophoran + tardigrade + arthropod) rather than arthropod lineage and that nuclear, lamin-derived proteins instead acquired new cytoplasmic roles at least three times independently in collembolans, copepods, and tardigrades. Transcriptomic and genomic data revealed three IF protein genes in the tardigrade Hypsibius dujardini, one of which (cytotardin) occurs exclusively in the cytoplasm of epidermal and foregut epithelia, where it forms belt-like filaments around each epithelial cell. These results suggest that a lamin derivative has been co-opted to enhance tissue stability in tardigrades, a function otherwise served by cytoplasmic IF proteins in all other bilaterians.

No MeSH data available.