Limits...
Distinct partitioning of ALS associated TDP-43, FUS and SOD1 mutants into cellular inclusions.

Farrawell NE, Lambert-Smith IA, Warraich ST, Blair IP, Saunders DN, Hatters DM, Yerbury JJ - Sci Rep (2015)

Bottom Line: Here we show that FUS variably partitioned to IPOD, JUNQ or alternate structures, contain a mobile fraction, were not microtubule dependent and initially did not contain ubiquitin.TDP-43 inclusions formed in a microtubule independent manner, did not contain a mobile fraction but variably colocalized to JUNQ inclusions and another alternate structure.We conclude that the RNA binding proteins TDP-43 and FUS do not consistently fit the currently characterised inclusion models suggesting that cells have a larger repertoire for generating inclusions than currently thought, and imply that toxicity in ALS does not stem from a particular aggregation process or aggregate structure.

View Article: PubMed Central - PubMed

Affiliation: Illawarra Health and Medical Research Institute, Wollongong, NSW 2522 Australia.

ABSTRACT
Amyotrophic lateral sclerosis is a rapidly progressing neurodegenerative disease associated with protein misfolding and aggregation. Most cases are characterized by TDP-43 positive inclusions, while a minority of familial ALS cases are instead FUS and SOD1 positive respectively. Cells can generate inclusions of variable type including previously characterized aggresomes, IPOD or JUNQ structures depending on the misfolded protein. SOD1 invariably forms JUNQ inclusions but it remains unclear whether other ALS protein aggregates arise as one of these previously described inclusion types or form unique structures. Here we show that FUS variably partitioned to IPOD, JUNQ or alternate structures, contain a mobile fraction, were not microtubule dependent and initially did not contain ubiquitin. TDP-43 inclusions formed in a microtubule independent manner, did not contain a mobile fraction but variably colocalized to JUNQ inclusions and another alternate structure. We conclude that the RNA binding proteins TDP-43 and FUS do not consistently fit the currently characterised inclusion models suggesting that cells have a larger repertoire for generating inclusions than currently thought, and imply that toxicity in ALS does not stem from a particular aggregation process or aggregate structure.

No MeSH data available.


Related in: MedlinePlus

Triple transfection reveals 3 distinct inclusions.NSC-34 cells were cotransfected with combinations of SOD1, TDP-43 and Htt including a triple transfection of all three plasmids and imaged after 48 h.
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f6: Triple transfection reveals 3 distinct inclusions.NSC-34 cells were cotransfected with combinations of SOD1, TDP-43 and Htt including a triple transfection of all three plasmids and imaged after 48 h.

Mentions: Given the different structural properties of TDP-43 and SOD1 inclusions, but their partial overlap when co-expressed, we performed triple transfections of TDP-43M337V-tomato, SOD1A4V -GFP and Httex146Q-Cerulean fluorescent protein. In this scenario, TDP-43 co-localized with SOD1A4V aggregates in some instances, but never co-localized with Httex146Q-Cerulean inclusions (Fig. 6). In some cases the cells were able to compartmentalize all three inclusions from one another. In these cases many of the TDP-43 inclusions appeared to be in the nucleus or very close to the nuclear envelope (Supplementary Material).


Distinct partitioning of ALS associated TDP-43, FUS and SOD1 mutants into cellular inclusions.

Farrawell NE, Lambert-Smith IA, Warraich ST, Blair IP, Saunders DN, Hatters DM, Yerbury JJ - Sci Rep (2015)

Triple transfection reveals 3 distinct inclusions.NSC-34 cells were cotransfected with combinations of SOD1, TDP-43 and Htt including a triple transfection of all three plasmids and imaged after 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4544019&req=5

f6: Triple transfection reveals 3 distinct inclusions.NSC-34 cells were cotransfected with combinations of SOD1, TDP-43 and Htt including a triple transfection of all three plasmids and imaged after 48 h.
Mentions: Given the different structural properties of TDP-43 and SOD1 inclusions, but their partial overlap when co-expressed, we performed triple transfections of TDP-43M337V-tomato, SOD1A4V -GFP and Httex146Q-Cerulean fluorescent protein. In this scenario, TDP-43 co-localized with SOD1A4V aggregates in some instances, but never co-localized with Httex146Q-Cerulean inclusions (Fig. 6). In some cases the cells were able to compartmentalize all three inclusions from one another. In these cases many of the TDP-43 inclusions appeared to be in the nucleus or very close to the nuclear envelope (Supplementary Material).

Bottom Line: Here we show that FUS variably partitioned to IPOD, JUNQ or alternate structures, contain a mobile fraction, were not microtubule dependent and initially did not contain ubiquitin.TDP-43 inclusions formed in a microtubule independent manner, did not contain a mobile fraction but variably colocalized to JUNQ inclusions and another alternate structure.We conclude that the RNA binding proteins TDP-43 and FUS do not consistently fit the currently characterised inclusion models suggesting that cells have a larger repertoire for generating inclusions than currently thought, and imply that toxicity in ALS does not stem from a particular aggregation process or aggregate structure.

View Article: PubMed Central - PubMed

Affiliation: Illawarra Health and Medical Research Institute, Wollongong, NSW 2522 Australia.

ABSTRACT
Amyotrophic lateral sclerosis is a rapidly progressing neurodegenerative disease associated with protein misfolding and aggregation. Most cases are characterized by TDP-43 positive inclusions, while a minority of familial ALS cases are instead FUS and SOD1 positive respectively. Cells can generate inclusions of variable type including previously characterized aggresomes, IPOD or JUNQ structures depending on the misfolded protein. SOD1 invariably forms JUNQ inclusions but it remains unclear whether other ALS protein aggregates arise as one of these previously described inclusion types or form unique structures. Here we show that FUS variably partitioned to IPOD, JUNQ or alternate structures, contain a mobile fraction, were not microtubule dependent and initially did not contain ubiquitin. TDP-43 inclusions formed in a microtubule independent manner, did not contain a mobile fraction but variably colocalized to JUNQ inclusions and another alternate structure. We conclude that the RNA binding proteins TDP-43 and FUS do not consistently fit the currently characterised inclusion models suggesting that cells have a larger repertoire for generating inclusions than currently thought, and imply that toxicity in ALS does not stem from a particular aggregation process or aggregate structure.

No MeSH data available.


Related in: MedlinePlus