DNA adducts of decarbamoyl mitomycin C efficiently kill cells without wild-type p53 resulting from proteasome-mediated degradation of checkpoint protein 1.
The mitomycin derivative 10-decarbamoyl mitomycin C (DMC) more rapidly activates a p53-independent cell death pathway than mitomycin C (MC). We recently documented that an increased proportion of mitosene1-beta-adduct formation occurs in human cells treated with DMC in comparison to those treated with MC. Here, we compare the cellular and molecular response of human cancer cells treated with MC and DMC. We find the increase in mitosene 1-beta-adduct formation correlates with a condensed nuclear morphology and increased cytotoxicity in human cancer cells with or without p53. DMC caused more DNA damage than MC in the nuclear and mitochondrial genomes. Checkpoint 1 protein (Chk1) was depleted following DMC, and the depletion of Chk1 by DMC was achieved through the ubiquitin proteasome pathway since chemical inhibition of the proteasome protected against Chk1 depletion. Gene silencing of Chk1 by siRNA increased the cytotoxicity of MC. DMC treatment caused a decrease in the level of total ubiquitinated proteins without increasing proteasome activity, suggesting that DMC mediated DNA adducts facilitate signal transduction to a pathway targeting cellular proteins for proteolysis. Thus, the mitosene-1-beta stereoisomeric DNA adducts produced by the DMC signal for a p53-independent mode of cell death correlated with reduced nuclear size, persistent DNA damage, increased ubiquitin proteolysis and reduced Chk1 protein.
Department of Biological Sciences, Hunter College and The Graduate Center, City University of New York, 695 Park Avenue, New York, NY 10065, USA.
Ernest K
EK
Boamah
Angelika
A
Brekman
Maria
M
Tomasz
Natura
N
Myeku
Maria
M
Figueiredo-Pereira
Senyene
S
Hunter
Joel
J
Meyer
Rahul C
RC
Bhosle
Jill
J
Bargonetti
D017209:Apoptosis; D045744:Cell Line, Tumor; D000071877:Checkpoint Kinase 1; D018736:DNA Adducts / Q000737:chemistry*; D004249:DNA Damage; D020868:Gene Silencing; D006801:Humans; D016685:Mitomycin / Q000633:toxicity; D008937:Mitomycins / Q000737:chemistry* / Q000633:toxicity; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
D018736:DNA Adducts; D008937:Mitomycins; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; C067795:10-decarbamoylmitomycin C; D016685:Mitomycin; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1; D046988:Proteasome Endopeptidase Complex