title: Essential role of T4 phage deoxycytidylate hydroxymethylase in a multienzyme complex for deoxyribonucleotide synthesis.
abstract: Several enzymes of deoxyribonucleoside triphosphate (dNTP) biosynthesis interact in T4 phage-infected Escherichia coli to form a multienzyme aggregate, the T4 dNTP synthetase complex. To test the specificity of enzyme interactions seen in vitro with this complex, we analyzed bacteria infected with four T4 gene 42 amber mutants, which specify truncated forms of dCMP hydroxymethylase, one of the constituent enzymes in the complex. Mutants that specify a nearly full length gene 42 product can form an intact complex, as revealed by two criteria: kinetic coupling among constituent enzymes in crude extracts of infected bacteria, and co-elution of enzyme activities from a gel filtration column. By these criteria, mutations that specify truncated proteins less than half the size of the full length protein cause disruption of the complex. These findings suggest that an enzymatically inactive form of dCMP hydroxymethylase can contribute toward assembly of an intact complex, so long as the incomplete protein is of sufficient size to fold normally, allowing interaction with other proteins in the complex.
authors.affiliation: Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.
authors.forename: C
authors.initials: C
authors.lastname: Thylén
authors.forename: C K
authors.initials: CK
authors.lastname: Mathews
mesh_terms: D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003854:Deoxyribonucleotides / Q000096:biosynthesis*; D004926:Escherichia coli / Q000201:enzymology*; D019877:Hydroxymethyl and Formyl Transferases; D007700:Kinetics; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D013604:T-Phages / Q000201:enzymology*; D014166:Transferases / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D003854:Deoxyribonucleotides; D009097:Multienzyme Complexes; D014166:Transferases; D019877:Hydroxymethyl and Formyl Transferases; C100226:deoxycytidylate hydroxymethyltransferase