Expression of the cDNA for the beta subunit of human casein kinase II confers partial UV resistance on xeroderma pigmentosum cells

Tal Teitz, Dalia Eli, Michal Penner, Mary Bakhanashvili, Tova Naiman, Terry L. Timme, Cada M. Wood, Robb Moses, Dan Canaani

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

An immortalized xeroderma pigmentosum cell line belonging to the complementation group D (XP-D) was transfected with a normal human cDNA clone library constructed in a mammalian expression vector. Following UV-irradiation-selection, a transformant having a stable, partially UV-resistant phenotype was isolated. A transfected cDNA of partial length was rescued from the transformant's cellular DNA by in vitro amplification, using expression-vector specific oligonucleotides as primers in a polymerase chain reaction (PCR). Expression of this cDNA complemented the UV sensitivity of the XP-D cell line to the UV-resistance levels characteristic of the primary transformant. The nucleotide sequence of the cDNA was determined. The deduced protein identified the cDNA as encoding for the beta subunit of casein kinase II (CKII-β). Similar to the effect exerted by the truncated CKII-β cDNA, expression of a cDNA clone encompassing the complete translated region of CKII-β leads to XP-D cells partially resistant to UV-irradiation. However, transfection of CKII-β cDNA could also partially complement the UV-sensitivity of a xeroderma pigmentosum cell line belonging to group C (XP-C). Analysis by Southern, Northern and RNAase mismatch cleavage techniques did not reveal any functional defect in the CKII-β gene of cell lines derived from either 7 XP-D or 10 XP-C families. We therefore consider it unlikely that either the XP-D or the XP-C DNA repair deficiency is associated with a defect in the beta subunit of casein kinase II. Nevertheless, our findings suggest the possibility that the cell's response to DNA damage is modulated by CKII-dependent protein phosphorylation.

Original languageEnglish (US)
Pages (from-to)85-97
Number of pages13
JournalMutation Research-DNA Repair
Volume236
Issue number1
DOIs
StatePublished - 1990
Externally publishedYes

Fingerprint

Casein Kinase II
Xeroderma Pigmentosum
Complementary DNA
Cells
Cell Line
Somatostatin-Secreting Cells
Clone Cells
DNA
DNA Repair-Deficiency Disorders
Irradiation
DNA Primers
Defects
Phosphorylation
Gene Library
Polymerase chain reaction
DNA Damage
Transfection
Oligonucleotides
Proteins
Amplification

Keywords

  • Casein kinase II
  • cDNA expression
  • Cell-cycle regulation
  • DNA
  • UV survival
  • Xeroderma pigmentosum

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Toxicology
  • Medicine(all)

Cite this

Expression of the cDNA for the beta subunit of human casein kinase II confers partial UV resistance on xeroderma pigmentosum cells. / Teitz, Tal; Eli, Dalia; Penner, Michal; Bakhanashvili, Mary; Naiman, Tova; Timme, Terry L.; Wood, Cada M.; Moses, Robb; Canaani, Dan.

In: Mutation Research-DNA Repair, Vol. 236, No. 1, 1990, p. 85-97.

Research output: Contribution to journalArticle

Teitz, Tal ; Eli, Dalia ; Penner, Michal ; Bakhanashvili, Mary ; Naiman, Tova ; Timme, Terry L. ; Wood, Cada M. ; Moses, Robb ; Canaani, Dan. / Expression of the cDNA for the beta subunit of human casein kinase II confers partial UV resistance on xeroderma pigmentosum cells. In: Mutation Research-DNA Repair. 1990 ; Vol. 236, No. 1. pp. 85-97.
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abstract = "An immortalized xeroderma pigmentosum cell line belonging to the complementation group D (XP-D) was transfected with a normal human cDNA clone library constructed in a mammalian expression vector. Following UV-irradiation-selection, a transformant having a stable, partially UV-resistant phenotype was isolated. A transfected cDNA of partial length was rescued from the transformant's cellular DNA by in vitro amplification, using expression-vector specific oligonucleotides as primers in a polymerase chain reaction (PCR). Expression of this cDNA complemented the UV sensitivity of the XP-D cell line to the UV-resistance levels characteristic of the primary transformant. The nucleotide sequence of the cDNA was determined. The deduced protein identified the cDNA as encoding for the beta subunit of casein kinase II (CKII-β). Similar to the effect exerted by the truncated CKII-β cDNA, expression of a cDNA clone encompassing the complete translated region of CKII-β leads to XP-D cells partially resistant to UV-irradiation. However, transfection of CKII-β cDNA could also partially complement the UV-sensitivity of a xeroderma pigmentosum cell line belonging to group C (XP-C). Analysis by Southern, Northern and RNAase mismatch cleavage techniques did not reveal any functional defect in the CKII-β gene of cell lines derived from either 7 XP-D or 10 XP-C families. We therefore consider it unlikely that either the XP-D or the XP-C DNA repair deficiency is associated with a defect in the beta subunit of casein kinase II. Nevertheless, our findings suggest the possibility that the cell's response to DNA damage is modulated by CKII-dependent protein phosphorylation.",
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