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Related Experiment Videos

Polymorphic sequences encoding the first open reading frame protein from LINE-1 ribonucleoprotein particles

V O Kolosha1, S L Martin

  • 1Department of Cellular and Structural Biology, University of Colorado School of Medicine, Denver 80262.

The Journal of Biological Chemistry
|February 10, 1995
PubMed
Summary
This summary is machine-generated.

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Mouse LINE-1 retrotransposons express multiple ORF1 proteins in F9 cells. These proteins are translated from L1 RNA enriched in ribonucleoprotein particles, indicating selective expression.

Area of Science:

  • Molecular Biology
  • Genomics
  • Retrotransposon Biology

Background:

  • LINE-1 (L1) retrotransposons are mobile genetic elements in mammalian genomes.
  • L1 elements possess two open reading frames (ORFs), ORF1 and ORF2.
  • Understanding L1 expression is crucial for studying genome dynamics and evolution.

Purpose of the Study:

  • To investigate the expression of the first open reading frame (ORF1) of mouse LINE-1 retrotransposons.
  • To characterize the different protein products encoded by ORF1.
  • To determine the cellular localization and enrichment of L1 RNA.

Main Methods:

  • Utilized mouse embryonal carcinoma cell line F9.
  • Isolated ribonucleoprotein particles (RNPs) and extracted RNA.

Related Experiment Videos

  • Employed reverse transcription-polymerase chain reaction (RT-PCR) to clone ORF1 regions.
  • Performed in vitro transcription and translation assays to examine coding capacity.

    • Main Results:

    • Identified three classes of ORF1 proteins with apparent molecular sizes of 41.3, 43, and 43.5 kDa.
    • Confirmed in vitro translation of 41.3 and 43 kDa proteins from full-length L1 RNA.
    • Recovered multiple ORF1 encoding sequences, suggesting expression from various L1 loci.
    • Observed selective enrichment of L1 sequences with intact ORF1 in RNPs.

    Conclusions:

    • Mouse F9 cells express multiple LINE-1 ORF1 protein variants.
    • L1 RNA is selectively packaged into ribonucleoprotein particles.
    • This suggests a regulated mechanism for LINE-1 expression and retrotransposition.