PMC:3654953 / 17465-19258
Annnotations
{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3654953","sourcedb":"PMC","sourceid":"3654953","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3654953","text":"GFAP is an intermediate filament (IF) protein expressed mainly by astrocytes and ependymocytes. Recent data suggested that GFAP-ϵ was unable to form filaments by itself but it could participate to the formation of the GFAP network by interacting with GFAP-α [20]. Hence we analyzed the IF meshwork in human astrocytoma U251-MG cells, constitutively expressing both GFAP-α and GFAP-ϵ, by expressing GFP-tagged wt and mutated GFAP-ϵ (GFP-GFAP-ϵwt vs. GFP-GFAP-ϵR430H). Cells were assigned to three patterns: [14] (i) exclusively filamentous pattern (F), (ii) cytoplasmic aggregates on a filamentous pattern (F + A), (iii) cytoplasmic aggregates with no filamentous pattern (A). The expression of GFP-GFAP-ϵwt led to a distribution among the three groups similar to that reported for GFP-GFAP-αwt[14] (Figure 2C) indicating no intrinsic damaging effect of recombinant GFP-GFAP-ϵwt in our experimental conditions. Contrariwise, expression of mutant GFP-GFAP-ϵR430H produced significant decrease in F (43% vs. 58%; test t p = 0.002) and increase in A (22% vs. 15%; test t p = 0.009) cells (Figure 2C), with a distinct distribution in the three patterns compared to GFP-GFAP-ϵwt expressing cells (ANOVA test for interaction p = 0.001). Notably, the expression of GFP-tagged GFAP carrying the R430C variant (GFP-GFAP-ϵR430C) led to a distribution amongst the three different patterns similar to that obtained with GFP-GFAP-ϵwt, i.e. non-significant (ANOVA test for interaction p = 0.333). These results indicate that GFAP-ϵR430H is inefficiently incorporated, and is likely to perturb the GFAP network in GFAP-expressing astrocytoma cells, whereas the GFAP-ϵR430C variant is functionally wt, but we cannot exclude the possibility that variations in the level of expression contributed to this result.","tracks":[{"project":"AxD_symptoms","denotations":[{"id":"T57","span":{"begin":307,"end":318},"obj":"Phenotype"},{"id":"T58","span":{"begin":1614,"end":1625},"obj":"Phenotype"}],"attributes":[{"id":"A57","pred":"hp_id","subj":"T57","obj":"http://purl.obolibrary.org/obo/HP_0009592"},{"id":"A58","pred":"hp_id","subj":"T58","obj":"http://purl.obolibrary.org/obo/HP_0009592"},{"subj":"T57","pred":"source","obj":"AxD_symptoms"},{"subj":"T58","pred":"source","obj":"AxD_symptoms"}]},{"project":"2_test","denotations":[{"id":"23634874-22912745-81641085","span":{"begin":259,"end":261},"obj":"22912745"},{"id":"23634874-18197187-81641086","span":{"begin":507,"end":509},"obj":"18197187"},{"id":"23634874-18197187-81641087","span":{"begin":794,"end":796},"obj":"18197187"},{"id":"T93218","span":{"begin":259,"end":261},"obj":"22912745"},{"id":"T91124","span":{"begin":507,"end":509},"obj":"18197187"},{"id":"T43725","span":{"begin":794,"end":796},"obj":"18197187"}],"attributes":[{"subj":"23634874-22912745-81641085","pred":"source","obj":"2_test"},{"subj":"23634874-18197187-81641086","pred":"source","obj":"2_test"},{"subj":"23634874-18197187-81641087","pred":"source","obj":"2_test"},{"subj":"T93218","pred":"source","obj":"2_test"},{"subj":"T91124","pred":"source","obj":"2_test"},{"subj":"T43725","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"AxD_symptoms","color":"#93d1ec","default":true},{"id":"2_test","color":"#ecec93"}]}]}}