Appendix B: Metabolic inhibitors used in virology

Most inhibitors are highly toxic, and it is essential that they are used strictly in accordance with all relevant safety instructions and procedures.

inhibitor

Molecular mass

Effect

Typical concentration for use

Cycloheximide (3-[2-{3,5-dimethyl- 281.4 2-oxocyclohexyl}-2hyd roxyethyi]glutarimide Puromycin (3'-[a-amino-pmethoxycinnamamido]-3'-dec)xyN, N-dimethyladenosine Castanospermine (8~13indolizidin e- 1(x,613,7(x,81[3-tetrol) 1-Deoxymannojirimycin (1,5dideoxy-1,5-imino-D-mannitol) 1-Deoxynojirimycin (1,5-dideoxy1,5-imino-D-glucitol) Monensin 544.4

Inhibitor of eukaryotic protein synthesis at translational level; used to induce high levels of mRNA prior to removal Protein synthesis inhibitor, Disrupts polyribosomes Inhibits processing of highmannose oligosccharides to complex-type inhibits processing of highmannose oligosaccharides to complex-type Inhibits formation of highmannose oligosaccharides Monovalent ionophore: Multiple effects on glycosylation due to disruption of passage through the Golgi Inhibits processing of highmannose oligosaccharides Prevents N-linked glycosylation

100--360 ~M

200 ~M

189.2

530-1320 ~M

199.6

1000-4000 ~I,M

163.2 692.9

1000-5000 ~I,M 1.0-1.5 ~M

Swainosine (8(x,13-indolizidinelc~,2(z,813-triol) Tunicamycin

173.2 840 approximately; very variable, multiple isomers 1255.4

O. 1-29 ~M 1.0-6 I~M

Actinomycin D

Irreversible inhibitor of transcription 0.4-8 IJM from double-standard DNA (RNA synthesis is inhibited at lower concentrations than is DNA synthesis)

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Inhibitor

Molecular mass

Effect

Typical concentration for use

Aphidicolin

338.5

Reversible inhibitor of eukaryotic 3-20 ~tM DNA polymerases cc and t3 and some viral DNA polymerases. May be used to synchronize cell growth Inhibits some viral DNA polymerases (notably in herpesviruses, where it is used to assay DNA synthesis-dependent protein synthesis) Inhibits RNA polymerases II (mRNA) and III (tRNA) 2140 #M

Phosphonoacetic acid

140.0

(z-Amanitin

919.0

0.001-0.01 I~M

(polymerase II) 10-100 #M (polymerase III) 200 ~tM

Cordycepin (3'-deoxyadenosine)
,...., m . .

251.2
..|..|..

Inhibits RNA synthesis and mRNA polyadenylation

9 9 .

This table is derived from Virology Labfax, D.R. Harper (Ed.), 1993, Bios Scientific Publishers, and is presented here with permission of the publishers.

364

Appendix B