Search citation statements
Paper Sections
Citation Types
Publication Types
Relationship
Authors
Journals
In Fanconi anemia (FA) cells the duration of the G2 phase of the cell cycle prolonged. Such a slowing of the G2 phase can be induced in normal cells by irradiation with gamma rays during S phase, which also further increases the duration of G2 in FA cells. The addition of caffeine during the last 7 h of culture shortens the G2 phase in both nonirradiated and irradiated FA cells. In nonirradiated normal cells it may have no effect or may increase G2 phase duration, but in irradiated normal reduces the slowing of G2 induced by the radiation. This suggests that FA cells recognize and repair preexisting DNA lesions during G2 phase and that caffeine inhibits this process. The principal anomaly in FA may be a deficient repair during S phase, as manifest in the prolonged postreplication repair period during G2 phase required to repair the larger number of lesions passing through S phase.
In Fanconi anemia (FA) cells the duration of the G2 phase of the cell cycle prolonged. Such a slowing of the G2 phase can be induced in normal cells by irradiation with gamma rays during S phase, which also further increases the duration of G2 in FA cells. The addition of caffeine during the last 7 h of culture shortens the G2 phase in both nonirradiated and irradiated FA cells. In nonirradiated normal cells it may have no effect or may increase G2 phase duration, but in irradiated normal reduces the slowing of G2 induced by the radiation. This suggests that FA cells recognize and repair preexisting DNA lesions during G2 phase and that caffeine inhibits this process. The principal anomaly in FA may be a deficient repair during S phase, as manifest in the prolonged postreplication repair period during G2 phase required to repair the larger number of lesions passing through S phase.
cDNAs encoding poly(ADP-ribose) polymerase from a human hepatoma lambda gt11 cDNA library were isolated by immunological screening. One insert of 1.3 kilobases (kb) consistently hybridized on RNA gel blots to an mRNA species of 3.6-3.7 kb, which is consistent with the size of RNA necessary to code for the polymerase protein (116 kDa). This insert was subsequently used in both in vitro hybrid selection and hybrid-arrested translation studies. An mRNA species from HeLa cells of 3.6-3.7 kb was selected that was translated into a 116-kDa protein, which was selectively immunoprecipitated with anti-poly (ADP-ribose) polymerase. To confirm that the 1.3-kb insert from lambda gt11 encodes for poly(ADP-ribose) polymerase, the insert was used to screen a 3- to 4-kb subset of a transformed human fibroblast cDNA library in the Okayama-Berg vector. One of these vectors [pcD-p(ADPR)P; 3.6 kb] was tested in transient transfection experiments in COS cells. This cDNA insert contained the complete coding sequence for polymerase as indicated by the following criteria: A 3-fold increase in in vitro activity was noted in extracts from transfected cells compared to mock or pSV2-CAT transfected cells. A 6-fold increase in polymerase activity in pcD-p(ADPR)P transfected cell extracts compared to controls was observed by "activity gel" analysis on gels of electrophoretically separated proteins at 116 kDa. A 10- to 15-fold increase in newly synthesized polymerase was detected by immunoprecipitation of labeled transfected cell extracts. Using pcD-p(ADPR)P as probe, it was observed that the level of poly(ADP-ribose) polymerase mRNA was elevated at 5 and 7 hr of S phase of the HeLa cell cycle, but was unaltered when artificial DNA strand breaks are introduced in HeLa cells by alkylating agents.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.