Structural details of the interaction
between pTyr1-CTD and the elongation factor Spt6
Pavel Brázda1, Magdaléna
Krejčíková1, Eliška Šmiřáková, Jiří Nováček1, Karel
Kubíček1, Patrick Cramer2 and Richard Štefl1
1CEITEC
– Central European Institute of Technology and Faculty of Science, Masaryk
University, CZ-62500 Brno, Czech Republic
2 Gene
Center and Department of Biochemistry, Ludwig-Maximilians-Universität München,
Feodor-Lynen-Strasse 25, D-81377 Munich, Germany
Post-transcriptional
modifications of the consensus motif
Y1-S2-P3-T4-S5-P6-S7 of the
C-terminal domain (CTD) of RNA polymerase II (RNAPII) are since last decade known as “CTD code” (1).
These modifications are dynamic and specific
for each phase of the transcriptional cycle (2). However,
how does the CTD code control the recruitment,
(de)activation, and displacement of relevant
processing factors, remains still poorly understood. Increased phosphorylation level of Y1 of
the CTD during the early elongation inhibits binding of termination factors, yet it stimulates binding of the elongation factor Spt6 through
its tandem SH2 (tSH2) domain (3). The limited structure information for the recognition of phospho-tyrosine modified CTD
that is currently available prevents deciphering of
how this important CTD modification mark is read out
by transcription factors (4,5). We set out to investigate this recognition
phenomenon using integrative approach to structural biology. Here,
we will show our combined
structural data from NMR, X-ray, and SAXS along with binding and biochemical assays on the study of recognition
Y1-phosphorylated CTD by the tSH2 domain of the elongation
factor Spt6.
1. Buratowski, S.:, The CTD
code. Nat Struct Biol., 10:679-80 (2003).
2. Egloff, S. and Murphy, S., Cracking
the RNA polymerase II CTD code, Trends Genet., 24: 280-8 (2008).
3. Mayer, A., Heidemann, M.,
Lidschreiber, M., Schreieck, Sun, M., Hintermair, C., Kremmer, E., Eick, D. and
Cramer, P., CTD Tyrosine Phosphorylation Impairs Termination Factor Recruitment
to RNA Polymerase II, Science, 336:1723-5 (2012).
4. Sun, M., Lariviere, L.,
Dengl, S., Mayer, A., Cramer, P., A tandem SH2 domain in transcription
elongation factor Spt6 binds the phosphorylated RNA polymerase II C-terminal
repeat domain (CTD), J.Biol.Chem. 285: 41597-41603 (2010).
5. Diebold, M., Loeliger, E.,
Koch, M., Winston, F., Cavarelli, J., Romier, C., Noncanonical Tandem Sh2
Enables Interaction of Elongation Factor Spt6 with RNA Polymerase II, J.Biol.Chem.
285: 38389 (2010).