Da. Bushnell et al., A MINIMAL SET OF RNA-POLYMERASE-II TRANSCRIPTION PROTEIN INTERACTIONS, The Journal of biological chemistry, 271(33), 1996, pp. 20170-20174
All pairwise interactions of RNA polymerase II and general transcripti
on factors (TF) IIB, E, F, and H have been quantitated by surface plas
mon resonance with the use of a Ni2+ chelate on the sensor surface whe
re necessary to attain higher sensitivity. Only 4 of 10 possible inter
actions were found above the detection limit: TFIIB, -E, and -F bindin
g RNA polymerase II and TFIIE binding to TFIIH. These four interaction
s constitute a minimal set for the formation of a transcription initia
tion complex and may represent the primary interactions involved in as
sembly of the complex. Point mutations in TFIIB that alter the locatio
n of transcription start sites in vivo markedly diminished the affinit
y of TFIIB binding to RNA polymerase II. Protein blotting revealed an
interaction between the largest subunit of TFIIE and third largest sub
unit of TFIIH, which may be responsible for the TFIIE binding to TFIIH
.