Skip to Content
Merck
  • Integrator restrains paraspeckles assembly by promoting isoform switching of the lncRNA NEAT1.

Integrator restrains paraspeckles assembly by promoting isoform switching of the lncRNA NEAT1.

Science advances (2020-09-15)
Jasmine Barra, Gabriel S Gaidosh, Ezra Blumenthal, Felipe Beckedorff, Mina M Tayari, Nina Kirstein, Tobias K Karakach, Torben Heick Jensen, Francis Impens, Kris Gevaert, Eleonora Leucci, Ramin Shiekhattar, Jean-Christophe Marine
ABSTRACT

RNA 3' end processing provides a source of transcriptome diversification which affects various (patho)-physiological processes. A prime example is the transcript isoform switch that leads to the read-through expression of the long non-coding RNA NEAT1_2, at the expense of the shorter polyadenylated transcript NEAT1_1. NEAT1_2 is required for assembly of paraspeckles (PS), nuclear bodies that protect cancer cells from oncogene-induced replication stress and chemotherapy. Searching for proteins that modulate this event, we identified factors involved in the 3' end processing of polyadenylated RNA and components of the Integrator complex. Perturbation experiments established that, by promoting the cleavage of NEAT1_2, Integrator forces NEAT1_2 to NEAT1_1 isoform switching and, thereby, restrains PS assembly. Consistently, low levels of Integrator subunits correlated with poorer prognosis of cancer patients exposed to chemotherapeutics. Our study establishes that Integrator regulates PS biogenesis and a link between Integrator, cancer biology, and chemosensitivity, which may be exploited therapeutically.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-PSPC1 antibody, Mouse monoclonal, clone 1L4, purified from hybridoma cell culture
Sigma-Aldrich
Insulin solution human, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Monoclonal Anti-Vinculin antibody produced in mouse, clone hVIN-1, ascites fluid
Sigma-Aldrich
Anti-INTS1 Antibody, clone 4.47, clone 4.47, from mouse
Sigma-Aldrich
Nutlin-3a, ≥98% (HPLC)
Sigma-Aldrich
Anti-INTS11 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Monoclonal Anti-PSF antibody produced in mouse, clone B92, purified from hybridoma cell culture
Sigma-Aldrich
Normal Rabbit IgG, Normal Rabbit IgG Polyclonal Antibody control validated for use in Immunoprecipitation & Western Blotting.