S. Spicuglia et al., TCR alpha enhancer activation occurs via a conformational change of a pre-assembled nucleoprotein complex, EMBO J, 19(9), 2000, pp. 2034-2045
The TCR alpha enhancer (E alpha) has served as a paradigm for studying how
enhancers organize trans-activators into nucleo-protein complexes thought t
o recruit and synergistically stimulate the transcriptional machinery. Litt
le is known, however, of either the extent or dynamics of E alpha occupancy
by nuclear factors during T cell development. Using dimethyl sulfate (DMS)
irt five footprinting, we demonstrate extensive Ea occupancy, encompassing
both previously identified and novel sites, not only in T cells representi
ng a developmental stage where E alpha is known to be active (CD4(+)CD8(+)-
DP cells), but surprisingly, also in cells at an earlier developmental stag
e where E alpha is not active (CD4(-)CD8(-)-DN cells). partial occupancy wa
s also established in B-lymphoid but not non-lymphoid cells. Irt vivo DNase
I footprinting, however, implied developmentally induced changes in nucleo
-protein complex topography. Stage-specific differences in factor compositi
on at E alpha sequences were also suggested by EMSA analysis. These results
, which indicate that alterations in the structure of a pre-assembled nucle
o-protein complex correlate with the onset of E alpha activity, may exempli
fy one mechanism by which enhancers can rapidly respond to incoming stimuli
.