The plant amino acid mimosine may inhibit initiation at origins of replication in Chinese hamster cells.
An understanding of replication initiation in mammalian cells has been hampered by the lack of mutations and/or inhibitors that arrest cells just prior to entry into the S period. The plant amino acid mimosine has recently been suggested to inhibit cells at a regulatory step in late G1. We have examined the effects of mimosine on cell cycle traverse in the mimosine [corrected]-resistant CHO cell line CHOC 400. When administered to cultures for 14 h after reversal of a G0 block, the drug appears to arrest the population at the G1/S boundary, and upon its removal cells enter the S phase in a synchronous wave. However, when methotrexate is administered to an actively dividing asynchronous culture, cells are arrested not only at the G1/S interface but also in early and middle S phase. Most interestingly, two-dimensional gel analysis of replication intermediates in the initiation locus of the amplified dihydrofolate reductase domain suggests that mimosine may actually inhibit initiation. Thus, this drug represents a new class of inhibitors that may open a window on regulatory events occurring at individual origins of replication.
Mosca
P J
PJ
Biochemistry Department, University of Virginia School of Medicine, Charlottesville 22908.
Dijkwel
P A
PA
Hamlin
J L
JL
D000818:Animals; D016590:Aphidicolin / Q000494:pharmacology; D016466:CHO Cells; D006224:Cricetinae; D004261:DNA Replication / Q000187:drug effects*; D015180:Electrophoresis, Gel, Two-Dimensional; D005434:Flow Cytometry; D016193:G1 Phase / Q000187:drug effects; D007700:Kinetics; D008898:Mimosine / Q000494:pharmacology*; D010944:Plants; D016192:Resting Phase, Cell Cycle / Q000187:drug effects; D016196:S Phase / Q000187:drug effects; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D013936:Thymidine / Q000037:antagonists & inhibitors
D016590:Aphidicolin; D008898:Mimosine; D013762:Tetrahydrofolate Dehydrogenase; D013936:Thymidine