Structural and dynamic changes of the serum response element and the core domain of serum response factor induced by their association.
Transcriptional activity of serum response factor (SRF) is dependent on its binding to the CC(A/T)(6)GG box (CArG box) of serum response element (SRE). By Raman spectroscopy, we carried out a comparative analysis, in solution, of the complexes obtained from the association of core-SRF with 20-mer SREs bearing wild-type and mutated c-fos CArG boxes. In case of association with the wild type c-fos CArG box, the complex does not bring out the expected Raman signature of a stable bending of the targeted SRE but keeps a bend-linear conformer oligonucleotide interconversion. The linear conformer population is larger than that of free oligonucleotide. In the core-SRF moiety of the wild-type complex a large spectral change associated with the CO-groups from Asp and/or Glu residues shows that their ionization states and the strength of their interactions decrease as compared to those of mutated non-specific complexes. Structural constraints evidenced on the free core-SRF are released in the wild-type complex and environmental heterogeneities appear in the vicinity of Tyr residues, due to higher water molecule access. The H-bonding configuration of one Tyr OH-group, in average, changes with a net transfer from H-bond acceptor character to a combined donor and acceptor character. A charge repartition distributed on both core-SRF and targeted SRE stabilizes the specific complex, allowing the two partners to experience a variety of conformations.
Stepánek
Josef
J
Laboratoire Acides Nucléiques & Biophotonique, FRE CNRS 3207, Université Pierre et Marie Curie, 5 rue Henri Desbruères, 91030 Evry, France.
Kopecký
Vladimír
V
Mezzetti
Alberto
A
Turpin
Pierre-Yves
PY
Paulin
Denise
D
Alpert
Bernard
B
Zentz
Christian
C
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D005786:Gene Expression Regulation; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D027602:Serum Response Element; D026362:Serum Response Factor / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013059:Spectrum Analysis, Raman; D014158:Transcription, Genetic; D014443:Tyrosine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D026362:Serum Response Factor; D014443:Tyrosine