title: Can terminators be used as insulators into yeast synthetic gene circuits?
abstract: BACKGROUND
  In bacteria, transcription units can be insulated by placing a terminator in front of a promoter. In this way promoter leakage due to the read-through from an upstream gene or RNA polymerase unspecific binding to the DNA is, in principle, removed. Differently from bacterial terminators, yeastS. cerevisiaeterminators contain a hexamer sequence, the efficiency element, that strongly resembles the eukaryotic TATA box i.e. the promoter sequence recognized and bound by RNA polymerase II.
  
  
  RESULTS
  By placing different yeast terminators (natural and synthetic) in front of the CYC1 yeast constitutive promoter stripped of every upstream activating sequences and TATA boxes, we verified that the efficiency element is able to bind RNA polymerase II, hence working as a TATA box. Moreover, terminators put in front of strong and medium-strength constitutive yeast promoters cause a non-negligible decrease in the promoter transcriptional activity.
  
  
  CONCLUSIONS
  Our data suggests that RNA polymerase II molecules upon binding the insulator efficiency element interfere with protein expression by competing either with activator proteins at the promoter enhancers or other RNA polymerase II molecules targeting the TATA box. Hence, it seems preferable to avoid the insulation of non-weak promoters when building synthetic gene circuit in yeastS. cerevisiae.
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nan Gang District, Harbin, 150080 People's Republic of China.
authors.forename: Wenjiang
authors.initials: W
authors.lastname: Song
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nan Gang District, Harbin, 150080 People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nan Gang District, Harbin, 150080 People's Republic of China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Liang
authors.affiliation: School of Life Science and Technology, Harbin Institute of Technology, 2 Yikuang Street, Nan Gang District, Harbin, 150080 People's Republic of China.
authors.forename: Mario Andrea
authors.initials: MA
authors.lastname: Marchisio
keywords: Efficiency element; Insulation; S. cerevisiae; TATA box; Terminator