Journal: Cell reports

Article Title: Mutant KRAS-driven selective mRNA translation reveals mechanisms and therapeutic vulnerabilities in cancer

doi: 10.1016/j.celrep.2026.117520

Figure Lengend Snippet: (A and B) Ribosome coverage on all ORFs showed increased ribosomes at the initiation site following sotorasib and MRTX1133 treatment in MiaPaca-2 and Panc10.05 cells, respectively ( p = 2.73 × 10 −2 ). No significant effect on the CDS in MRTX1133-treated Panc10.05 cells ( p = 0.414). (C) Ribosome coverage on TE down transcripts showed the accumulation of ribosomes upstream and downstream of the start site following sotorasib treatment in MiaPaca-2 ( p = 4.00 × 10 𢈒2 ), suggesting a slowdown in translation. (D) Ribosome coverage on TE down transcripts showed the accumulation of ribosomes upstream of the stop site following sotorasib treatment in MiaPaca-2 ( p = 1.66 × 10 −4 ), suggesting a block in translation elongation. (E) An increase of ribosome coverage between 70 and 90 nt region was observed in TE up transcripts following sotorasib (blue) treatment over control (black) in the 5′UTR and CDS around the start site ( p = 1.73 × 10 −2 ). (F) Fraction of pause-site codons (RxK, KKK, PP, PD, and PG) across the detected ORFs for the first 50 codons. (G, H, and I) Sucrose gradient fractionation of RNAse I digested polysome showing the accumulation of disome in MiaPaca-2 cells treated with sotorasib and quantification of the disome peak relative to DMSO ( p < 0.05). Immunoblot shows RPL5 and RPS10 in the indicated fractions collected from RNase I-digested polysome following DMSO vs. sotorasib treatment in MiaPaca-2. (J, K, and L) Immunoblot shows the induction of ZNF598, EDF1, p-ZAKα, ZAKα, RPS10-Ub following sotorasib treatment in MiaPaca-2. Anisomycin (ANS) treatment is included as a positive control for ribosome stalling and collision-induced stress signaling. β-actin is used as a loading control. (M) Mutant KRAS G12C inhibitor sotorasib affects translation initiation and elongation, resulting in ribosome stalling, while mutant KRAS G12D inhibitor MRTX1133 predominantly reduces translation initiation.

Article Snippet: The following primary antibodies were used as per the manufacturer’s recommended dilution: ZNF598 (Bethyl A305-108A), EDF1 (ProteinTech 12419-1-AP), ZAKα (Bethyl A301-993A), RPS6 (Cell Signaling Technology 2317), p-S6 (Cell Signaling Technology 2215), EIF4E (Cell Signaling Technology 9742), EEF1A1 (ProteinTech 11402-1-AP), EEF2 (Cell Signaling Technology 2332), p -ERK (Cell Signaling Technology 9101), ERK (Cell Signaling Technology 4695), RPL5 (Cell Signaling Technology 51345S), RPS10 (Abcam ab151550 and Invitrogen PA596460 ), DLAT (Cell Signaling Technology 12362), PDHB (Invitrogen MA5-32757), ECHS1 (ProteinTech 66117-1-LG), ACADM (ProteinTech 55210-1-AP), β-actin (Sigma-Aldrich A5316).

Techniques: Blocking Assay, Control, Fractionation, Western Blot, Positive Control, Mutagenesis

Journal: Molecular Cell

Article Title: Mechanism of RACK1-dependent ZAKα activation at stalled and collided ribosomes

doi: 10.1016/j.molcel.2026.04.034

Figure Lengend Snippet: Molecular dynamics simulation video of ZAKα-RACK1 corresponding to Figures 7F–7H and S7 K. ZAKα residues 336 to 417 (SAM domain, green) and 309 to 329 (C-terminal part of LZ, blue) and RACK1 (orange) are highlighted. Time frame, 100 ns.

Article Snippet: Antibodies used: Rabbit polyclonal anti-ZAKα (Bethyl, Cat#A301-993A; RRID: AB_1576612 ), Mouse monoclonal anti-phospho-p38 (Cell Signaling, Cat#9216; RRID: AB_331296 ) Rabbit monoclonal anti-phospho-p38 (Cell Signaling, Cat#4511S; RRID: AB_2139682 ), Rabbit polyclonal antibody anti-p38 (Cell Signaling, Cat#9212; RRID: AB_330713 ), Mouse monoclonal anti-phospho-SAPK/JNK (Cell Signaling, Cat#9255; RRID: AB_2307321 ), Rabbit monoclonal anti-phospho-SAPK/JNK (Cell Signaling, Cat#4668, RRID: AB_823588 ), Rabbit monoclonal anti-SAPK/JNK (Cell Signaling, Cat#9258; RRID: AB_2141027 ), Rabbit polyclonal anti-ZAK (Proteintech, Cat#14945-1-AP; RRID: AB_1064269 ), Mouse monoclonal anti-p150 (BD biosciences, Cat#610473; RRID: AB_397845 ), Mouse monoclonal anti-α-Tubulin (Merck, Cat#T9026; RRID: AB_477593 ), Mouse monoclonal anti-HA-tag (Santa Cruz Biotechnology, Cat#sc-7392 HRP; RRID: AB_2894930 ), Mouse monoclonal anti-Puromycin (Millipore, Cat#MABE343; RRID: AB_2566826 ), Rabbit polyclonal anti-RACK1 (Bethyl, Cat#A302-545A; RRID: AB_1999012 ), Rabbit monoclonal anti-RPS10 (Abcam # ab151550; RRID: AB_2714147 ), Rabbit polyclonal anti-GST (Sigma, Cat#G7781; RRID: AB_259965 ), Rabbit polyclonal anti-GCN1 (Thermo Fisher, A301-843A; RRID: AB_1264319 ), Rabbit polyclonal anti-GCN2 (Cell Signaling, Cat#3302S; RRID: AB_2277617 ), Mouse monoclonal anti-GFP (Roche Cat# 11814460001; RRID: AB_390913 ), Mouse monoclonal anti-gamma-tubulin (Sigma, #T5326; RRID: AB_532292 ), Mouse monoclonal anti-RPL19 (Novus Biologicals #H00006143-M01; RRID: AB_509253 ).

Techniques: