Structural study of the yeast RNA polymerase A. Electron microscopy of lipid-bound molecules and two-dimensional crystals.
Two-dimensional crystals of yeast RNA polymerase A (I) were obtained by interaction with positively charged lipid layers. The analysis of single molecular images of lipid-bound RNA polymerases showed that the enzyme was preferentially oriented by the lipid phase, which probably facilitated crystallization. Electron micrographs of the crystals revealed a rectangular unit cell 25.8 nm by 45.6 nm in size containing four RNA polymerase dimers related by P22(1)2(1) symmetry. The projection map showed, at about 2.5 nm resolution, two different views of the enzyme characterized by two bent arms, which appeared to cross at one end. These arms are likely to contain the A190 and A135 subunits and delimit a 3 to 4 nm wide groove. Additional structural features were observed and compared to the Escherichia coli enzyme.
Schultz
P
P
Laboratoire de Génétique Moléculaire des Eucaryotes, Strasbourg, France.
Célia
H
H
Riva
M
M
Darst
S A
SA
Colin
P
P
Kornberg
R D
RD
Sentenac
A
A
Oudet
P
P
D008051:Lipid Bilayers / Q000378:metabolism; D008854:Microscopy, Electron; D011485:Protein Binding; D011487:Protein Conformation; D012318:RNA Polymerase I / Q000378:metabolism / Q000648:ultrastructure*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D008051:Lipid Bilayers; D012318:RNA Polymerase I