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 language | English |
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Pages (from-to) | 85-97 |
Number of pages | 13 |
Journal | Mutation Research - DNA Repair |
Volume | 236 |
Issue number | 1 |
DOIs | |
State | Published - Jul 1990 |
Externally published | Yes |
Bibliographical note
Funding Information:We are grateful to Z. Lev and E. Yagil for a computer search of sequence homology. This research was supported by USPHS grants 37860 and
Funding Information:
AG07123 to R.E.M. and grants awarded to D.C. by the Fund for Basic Research administered by the Israel Academy of Sciences and Humanities, the United States-Israel Binational Science Foundation, the Hareli Fund for Cancer Research and a Research Career Development Award from the Israel Cancer Research Fund.
Funding
We are grateful to Z. Lev and E. Yagil for a computer search of sequence homology. This research was supported by USPHS grants 37860 and AG07123 to R.E.M. and grants awarded to D.C. by the Fund for Basic Research administered by the Israel Academy of Sciences and Humanities, the United States-Israel Binational Science Foundation, the Hareli Fund for Cancer Research and a Research Career Development Award from the Israel Cancer Research Fund.
Funders | Funder number |
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Hareli Fund for Cancer Research | |
National Institute on Aging | P01AG007123 |
Israel Cancer Research Fund | |
U.S. Public Health Service | 37860 |
Naman Family Fund for Basic Research | |
United States-Israel Binational Science Foundation | |
Israel Academy of Sciences and Humanities |
Keywords
- Casein kinase II
- Cell-cycle regulation
- DNA
- UV survival
- Xeroderma pigmentosum
- cDNA expression