Journal: Biochemical and biophysical research communications

Article Title: Hsp104 is essential for the selective degradation in yeast of polyglutamine expanded ataxin-1 but not most misfolded proteins generally

doi: 10.1016/j.bbrc.2009.12.018

Figure Lengend Snippet: Mutant human ataxin-1 with expanded polyglutamine repeats (82Q) is selectively degraded by the proteasome in yeast. Yeast cells (JN284) were transfected with the plasmids carrying human ataxin-1 (30Q or 82Q) whose expression is under the control of the GAL1 promoter. Cells were grown in the minimal medium containing 2% raffinose until mid-log phase (OD600 = 0.5–1.0) and then transferred to the induction medium containing 2% galactose for 6–8 h. The degradation of ataxin-1 in these cells was measured in the presence or absence of MG132 (50 µM) either by pulse-chase analysis and immunoprecipitation with the anti-ataxin-1 antibody 11750 or by the promoter shut-off assay. Similar results were obtained in three independent experiments.

Article Snippet: The yeast expression plasmids carrying human ataxin-1 (wild-type and mutant) under the control of GAL1 promoter – provided by Prof. Huda Zoghbi (Baylor College of Medicine) – were constructed by inserting human ATXN1 (SCA1) coding region into SpeI and SalI sites of p416 Gal1 or p423 Gal1 vector (ATCC).

Techniques: Mutagenesis, Transfection, Expressing, Control, Pulse Chase, Immunoprecipitation

Journal: Biochemical and biophysical research communications

Article Title: Hsp104 is essential for the selective degradation in yeast of polyglutamine expanded ataxin-1 but not most misfolded proteins generally

doi: 10.1016/j.bbrc.2009.12.018

Figure Lengend Snippet: Hsp104 is required for the selective degradation of mutant human ataxin-1. (A) Selective degradation of mutant ataxin-1 [82Q] does not require Ydj1p, an Hsp40 homolog, (B) but does require Hsp104. Plasmids carrying the mutant [82Q] ataxin-1 were transfected into (A) the wild-type (W303-1b) and temperature-sensitive mutant strain of Ydj1p (ACY 17b: ydj1–151) or (B) the wild-type (W303a) and Hsp104-deletion mutant strain (YS 483: Δhsp104). Degradation of ataxin-1 was then measured as described in Fig. 1. (C) Transfection of Hsp104 into the Hsp104-deletion strain restores the degradation of mutant ataxin-1 back to the control level. A CEN plasmid carrying the wild-type Hsp104 (under the GAL1 promoter) was introduced into Δhsp104 cells expressing the mutant ataxin-1, and its degradation was measured. (D) Hsp104 deletion does not affect the degradation of the model substrate ubiquitin-Pro-β-galactosidase (Ub-Pro-β-gal).

Article Snippet: The yeast expression plasmids carrying human ataxin-1 (wild-type and mutant) under the control of GAL1 promoter – provided by Prof. Huda Zoghbi (Baylor College of Medicine) – were constructed by inserting human ATXN1 (SCA1) coding region into SpeI and SalI sites of p416 Gal1 or p423 Gal1 vector (ATCC).

Techniques: Mutagenesis, Transfection, Control, Plasmid Preparation, Expressing, Ubiquitin Proteomics

Journal: iScience

Article Title: Niche-expressed Galectin-1 is involved in pre-B acute lymphoblastic leukemia relapse through pre-B cell receptor activation

doi: 10.1016/j.isci.2023.106385

Figure Lengend Snippet: GAL1+ mesenchymal stromal cells favor the development of murine Pre-BCR+ B-ALL

Article Snippet: Pre-B ALL from patient #IPC6 and Nalm6 were incubated 30 min at 4 °C in PBS with 70 nM recombinant human GAL1 (kind gift from Dr. L. Elantak) in absence or in presence of different concentrations of thiodigalactoside or of the GAL1 inhibitor OTX008 (MedChemExpress).

Techniques:

Journal: iScience

Article Title: Niche-expressed Galectin-1 is involved in pre-B acute lymphoblastic leukemia relapse through pre-B cell receptor activation

doi: 10.1016/j.isci.2023.106385

Figure Lengend Snippet: Pre-BCR signaling and human pre-B ALL growth are impaired in the absence of GAL1

Article Snippet: Pre-B ALL from patient #IPC6 and Nalm6 were incubated 30 min at 4 °C in PBS with 70 nM recombinant human GAL1 (kind gift from Dr. L. Elantak) in absence or in presence of different concentrations of thiodigalactoside or of the GAL1 inhibitor OTX008 (MedChemExpress).

Techniques: