Enzymatic phosphorylation of unnatural nucleosides

Y. Wu, M. Fa, E.L. Tae, P.G. Schultz, F.E. Romesberg, J. Am. Chem. Soc. (2002) 124:14626-14630.
pubpic2002wuWe report our initial efforts toward the development of an unnatural in vivo nucleoside phosphorylation pathway that is based on nucleoside salvage enzymes.


Excited state dynamics and heterogeneity of folded and unfolded states of cytochrome c

R. Jimenez, F.E. Romesberg, J. Phys. Chem. B. (2002) 106(35):9172-9180.
pubpic2002jimenezTransient absorption, transient grating, and 3PEPS measurements are used to characterize the photophysics of the heme chromophore in the folded protein and in two different unfolded proteins. We find that cyt c has little structural heterogeneity in the folded state, and a larger degree of structural heterogeneity in the unfolded states that depends on the unfolding conditions.


Previously uncharacterized genes in the UV- and MMS-induced DNA damage response in yeast

D. Hanway, J.K. Chin, G. Xia, G. Oshiro, E.A. Winzeler, F.E. Romesberg, Proc. Natl. Acad. Sci. USA (2002) 99:10605-10610.
pubpic2002hanwayA competitive growth assay is used to identify yeast genes involved in the repair of UV- or MMS-induced DNA damage using a collection of 2,827 yeast strains in which each strain has a single ORF replaced with a cassette containing two unique sequence tags. Identified genes include three uncharacterized ORFs, as well as genes that encode protein products implicated in ubiquitination, gene silencing, and transport across the mitochondrial membrane.


Directed evolution of novel polymerase activities: mutation of a DNA polymerase into and efficient RNA polymerase

G. Xia, L. Chen, T. Sera, M. Fa, P.G. Schultz, F.E. Romesberg, Proc. Natl. Acad. Sci. USA (2002) 99:6597-6602.
pubpic2002xiaWe use an activity-based phage display method to screen a Stoffel fragment (SF) polymerase library, and we identify three SF mutants that incorporate ribonucleoside triphosphates virtually as efficiently as the wild-type SF polymerase incorporates dNTP substrates.