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Related Experiment Videos

Cloning, expression and characterization of human tissue-specific DNA polymerase lambda2.

Fu Gu1, Chun You, JianPing Liu

  • 1State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China.

Science in China. Series C, Life Sciences
|July 27, 2007
PubMed
Summary
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A novel DNA polymerase lambda2 variant was identified, playing a role in DNA repair. Its aberrant expression in hepatocellular carcinoma suggests involvement in liver cancer development.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • DNA polymerase lambda is crucial for DNA repair and recombination.
  • A new variant, POL lambda2, was discovered in human liver cDNA.

Purpose of the Study:

  • To characterize the novel POL lambda2 variant.
  • To investigate its role in hepatocellular carcinoma (HCC).

Main Methods:

  • Cloning and sequencing of POL lambda2.
  • Bioinformatics analysis for gene structure and protein domains.
  • Cellular localization studies in transfected cells.
  • Expression analysis in human tissues and HCC specimens.
  • Recombinant protein expression, purification, and in vitro activity assays.

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Main Results:

  • POL lambda2 gene spans 7.9 kb on chromosome 10q24, with 8 exons and 7 introns.
  • It possesses DNA polymerase Xc and BRCT domains and localizes to the nucleus.
  • Abundant expression in liver and testis; weak in ovary; undetectable elsewhere.
  • Aberrant POL lambda/POL lambda2 expression ratio in 80% of HCC tissues due to POL lambda upregulation.
  • Purified recombinant POL lambda2 demonstrated DNA polymerase activity in vitro.

Conclusions:

  • The novel POL lambda2 variant has characteristic DNA polymerase domains and nuclear localization.
  • Upregulated POL lambda expression in HCC, altering the POL lambda/POL lambda2 ratio, may contribute to hepatocarcinogenesis.