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Novel RNA-Binding Proteins Isolation by the RaPID Methodology
11:19

Novel RNA-Binding Proteins Isolation by the RaPID Methodology

Published on: September 30, 2016

LRP130, a single-stranded DNA/RNA-binding protein, localizes at the outer nuclear and endoplasmic reticulum membrane,

Naoto Tsuchiya1, Hirokazu Fukuda, Katsuhiko Nakashima

  • 1Biochemistry Division, National Cancer Center Research Institute, 5-1-1 Tsukiji Chuo-ku, Tokyo 104-0045, Japan.

Biochemical and Biophysical Research Communications
|April 15, 2004
PubMed

Insights

Leucine-rich repeat-containing protein 130 (LRP130) binds RNA and is found in the nuclear envelope and endoplasmic reticulum. This protein may regulate mRNA metabolism at the nuclear envelope and ER.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Leucine-rich repeat-containing protein 130 (LRP130), also known as LRPPRC, is an RNA and single-stranded DNA-binding protein.
  • LRP130 is a candidate gene for Leigh syndrome, a French-Canadian type cytochrome c oxidase deficiency, but its biological function remains unclear.

Purpose of the Study:

  • To investigate the RNA-binding capabilities and subcellular localization of LRP130.
  • To elucidate the role of LRP130 in cellular processes, particularly mRNA metabolism.

Main Methods:

  • Binding assays with synthetic RNA homopolymers and a repetitive RNA sequence.
  • Subcellular fractionation of mouse and HeLa cells to isolate nuclear, endoplasmic reticulum (ER), and mitochondrial components.
  • Immunostaining of HeLa cells to visualize LRP130 localization.
  • Overexpression of EGFP-fused LRP130 to observe effects on poly(A)(+) RNA distribution.

Main Results:

  • The C-terminal region of mouse LRP130 (a.a. 845-964) binds to various RNA polymers.
  • LRP130 localizes to nuclear/ER and mitochondrial fractions, with specific enrichment at the outer nuclear membrane (ONM)/ER.
  • LRP130 associates with poly(A)(+) RNA at the ONM/ER.
  • Overexpression of LRP130 leads to nuclear accumulation of poly(A)(+) RNA.

Conclusions:

  • LRP130 is an RNA-binding protein with a significant role in mRNA metabolism.
  • LRP130 associates with mRNA/mRNP complexes at the outer nuclear envelope and ER.
  • LRP130 may control mRNA metabolism by interacting with these complexes at the nuclear periphery.

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