PMID- 14705778
OWN - NLM
STAT- MEDLINE
DCOM- 20040526
LR  - 20161124
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 322
IP  - 1
DP  - 2003 Nov 1
TI  - A novel Escherichia coli strain allows functional analysis of guanylate kinase 
      drug resistance and sensitivity.
PG  - 40-7
AB  - Guanylate kinase is a critical enzyme in the biosynthesis of guanosine 
      5'-triphosphate (GTP) and dGTP and is responsible for the phosphorylation of 
      guanosine 5'-monophosphate (GMP) and dGMP to guanosine 5'-diphosphate (GDP) and 
      dGDP, respectively. As with many nucleotide-metabolizing enzymes, guanylate 
      kinase is involved in antimicrobial and antineoplastic drug activation. This is 
      due to the structural similarities of such agents with nucleobases or nucleosides 
      that are acted upon by endogenous enzymes. Despite the involvement of guanylate 
      kinase in 6-thioguanine, mercaptopurine, and abasic guanosine analog (e.g., 
      ganciclovir) activation, studies have only recently focused on the molecular 
      basis of the structure to function relationship of a mammalian guanylate kinase. 
      As a means to evaluate the details of amino acid side chain involvement in 
      substrate interaction, we have constructed a conditional 
      guanylate-kinase-deficient Escherichia coli strain that requires the presence of 
      a functional, plasmid-borne guanylate kinase for growth under selective 
      conditions. Positive genetic selection provides a rapid mechanism to identify not 
      only functional guanylate kinase mutants but also those that result in drug 
      resistance. This novel strain will be beneficial to assess the role of specific 
      amino acids of guanylate kinase in structure, function, drug activation, and drug 
      resistance.
FAU - Stolworthy, Tiffany S
AU  - Stolworthy TS
AD  - Department of Pharmaceutical Sciences, P.O. Box 646534, Washington State 
      University, Pullman, WA 99164-6534, USA.
FAU - Krabbenhoft, Elizabeth
AU  - Krabbenhoft E
FAU - Black, Margaret E
AU  - Black ME
LA  - eng
GR  - R01 CA85939/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 85-32-5 (Guanosine Monophosphate)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - EC 2.7.4.4 (Nucleoside-Phosphate Kinase)
RN  - EC 2.7.4.8 (Guanylate Kinases)
RN  - FTK8U1GZNX (Thioguanine)
SB  - IM
MH  - Chromosomes, Bacterial
MH  - Cloning, Molecular
MH  - Drug Resistance, Microbial
MH  - Escherichia coli/enzymology/*genetics
MH  - Gene Silencing
MH  - Guanosine Monophosphate/metabolism
MH  - Guanosine Triphosphate/metabolism
MH  - Guanylate Kinases
MH  - Mutation
MH  - Nucleoside-Phosphate Kinase/deficiency/*metabolism
MH  - Phosphorylation
MH  - Plasmids
MH  - Thioguanine/pharmacology
EDAT- 2004/01/07 05:00
MHDA- 2004/05/27 05:00
CRDT- 2004/01/07 05:00
PHST- 2003/04/24 00:00 [received]
PHST- 2004/01/07 05:00 [pubmed]
PHST- 2004/05/27 05:00 [medline]
PHST- 2004/01/07 05:00 [entrez]
AID - S0003269703004883 [pii]
AID - 10.1016/j.ab.2003.07.005 [doi]
PST - ppublish
SO  - Anal Biochem. 2003 Nov 1;322(1):40-7. doi: 10.1016/j.ab.2003.07.005.