Expression and Characterization of the Human rpS3 in a Methylotrophic Yeast Pichia pastoris

A human ribosomal protein S3 (rpS3), which also functions as a DNA repair enzyme (UV endonuclease III), was expressed in a methylotrophic yeast, Pichia pastoris, and biochemically characterized. UV endonuclease activity was previously characterized, and this activity of mammalian rpS3 was found to be non-specific upon purification and storage. Under the Pichia expression system, the subcloned cDNA of the human rpS3 gene revealed a peptide of 42 kDa by SDS-PAGE and Western blot. The secreted form of human rpS3 rendered no endonuclease activity while the intracellular form showed UV specific endonuclease activity by the nick circle assay.

[1]  Lim Young-Hee.  Rapid PCR Method for Detecting Candida albicans Using Primers Derived from the Integrin-like Protein Gene αINT1 of Candida albicans , 2000 .

[2]  H. Youn,et al.  A Recombinant Mouse GM-CSF Protein Expressed as an Inclusion Form Shows Colony Stimulating Activity , 2000 .

[3]  Yong-Keel Choi,et al.  PCR-RFLP and Sequence Analysis of the rDNA ITS Region in the Fusarium spp. , 2000 .

[4]  J. Cregg,et al.  Heterologous protein expression in the methylotrophic yeast Pichia pastoris. , 2000, FEMS microbiology reviews.

[5]  L. Paulin,et al.  The complete genome sequence of PM2, the first lipid-containing bacterial virus To Be isolated. , 1999, Virology.

[6]  Jae Yung Lee,et al.  Study on the Specificity Alteration of Mammalian UV Endonuclease III , 1997 .

[7]  R. Fellous,et al.  Implication of Mammalian Ribosomal Protein S3 in the Processing of DNA Damage (*) , 1995, The Journal of Biological Chemistry.

[8]  William C. Raschke,et al.  Recent Advances in the Expression of Foreign Genes in Pichia pastoris , 1993, Bio/Technology.

[9]  J. Strathern,et al.  Recovery of plasmids from yeast into Escherichia coli: shuttle vectors. , 1991, Methods in enzymology.

[10]  J. Cregg,et al.  Structural comparison of the Pichia pastoris alcohol oxidase genes , 1989, Yeast.

[11]  S. Linn,et al.  Purification and characterization of UV endonucleases I and II from murine plasmacytoma cells. , 1989, Journal of Biological Chemistry.

[12]  K. Kraemer,et al.  DNA repair protects against cutaneous and internal neoplasia: evidence from xeroderma pigmentosum. , 1984, Carcinogenesis.

[13]  K. Murata,et al.  Transformation of intact yeast cells treated with alkali cations , 1983 .

[14]  S. Linn,et al.  An altered apurinic DNA endonuclease activity in group A and group D xeroderma pigmentosum fibroblasts. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[15]  R. Espejo,et al.  Properties and Characterization of the Host Bacterium of Bacteriophage PM2 , 1968, Journal of bacteriology.