Genetic characterization and isolation of theSaccharomyces cerevisiae gene coding for uridine monophosphokinase

Liljelund, Patricia; Lacroute, François

doi:10.1007/bf02428034

Cited by 48 publications

(27 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For dTDP synthesis, the reaction rate reaches maximum at pH 7 $ 8 then decline with further increasing pH. The results are similar to that of the enzyme reactions using the same enzyme but different substrates, nucleoside monophosphates (NMPs) such as AMP, GMP, CMP, and TMP Konrad, 1992Konrad, , 1993Liljelund and Lacroute, 1986;Ma et al, 1990). The optimal pH for the dNDP synthesis coincides with that of the pyruvate kinase catalyzed phosphorylation from dNDP to dNTP, the second step phosphorylation in the dNTP biosynthesis from dNMP .…”

Section: Analysis Of Amino Acid Sequence Of the Dnmp Kinasessupporting

confidence: 71%

“…The yeast GUK1 encoding GK has been cloned in E. coli in laboratory scale (Konrad, 1992;Li et al, 1996). The CK and TK have been purified and characterized from the wild S. cerevisiae strain Jong et al, , 1993Liljelund and Lacroute, 1986;Ma et al, 1990). In previous studies, these kinase enzymes are extracted from bacterial sources or animal tissues and used to study the physiological and metabolic functions in the living organisms (Yan and Tsai, 1999).…”

Section: Discussionmentioning

confidence: 99%

“…URA6 encoding both uridine monophosphate kinase and deoxycytidylate monophosphate kinase from S. cerevisiae has been purified and characterized from the wild type S. cerevisiae strain (Jong et al, 1993;Liljelund and Lacroute, 1986;Ma et al, 1990). The PCR product was cloned into the BamHI-EcoRI sites of pUC19 to give the pUC19-URA6 cloning plasmid and then transformed into E. coli strain DH5alpha.…”

Section: Cloning and Over-expression Of Dnmp Kinase Genesmentioning

confidence: 98%

See 2 more Smart Citations

Cloning of deoxynucleoside monophosphate kinase genes and biosynthesis of deoxynucleoside diphosphates

Bao

Ryu²

2006

Biotech & Bioengineering

View full text Add to dashboard Cite

The genes encoding four deoxynucleoside monophosphate kinase (dNMP kinase) enzymes, including ADK1 for deoxyadenylate monophosphate kinase (AK), GUK1 for deoxyguanylate monophosphate kinase (GK), URA6 for deoxycytidylate monophosphate kinase (CK), and CDC8 for deoxythymidylate monophosphate kinase (TK), were isolated from the genome of Saccharomyces cerevisiae ATCC 2610 strain and cloned into E. coli strain BL21(DE3). Four recombinant plasmids, pET17b-JB1 containing ADK1, pET17b-JB2 containing GUK1, pET17b-JB3 containing URA6, and pET17b-JB4 containing CDC8, were constructed and transformed into E. coli strain for over-expression of AK, GK, CK, and TK. The amino acid sequences of these enzymes were analyzed and a putative conserved peptide sequence for the ATP active site was proposed. The four deoxynucleoside diphosphates (dNDP) including deoxyadenosine diphosphate (dADP), deoxyguanosine diphosphate (dGDP), deoxycytidine diphosphate (dCDP), and deoxythymidine diphosphate (dTDP), were synthesized from the corresponding deoxynucleoside monophosphates (dNMP) using the purified AK, GK, CK, and TK, respectively. The effects of pH and magnesium ion concentration on the dNDP biosynthesis were found to be important. A kinetic model for the synthetic reactions of dNDP was developed based on the Bi-Bi random rapid equilibrium mechanism. The kinetic parameters including the maximum reaction velocity and Michaelis-Menten constants were experimentally determined. The study on dNDP biosynthesis reported in this article are important to the proposed bioprocess for production of deoxynucleoside triphosphates (dNTP) that are used as precursors for in vitro DNA synthesis. There is a significant advantage of using enzymatic biosyntheses of dNDP as compared to the chemical method that has been in commercial use.

show abstract

Section: Analysis Of Amino Acid Sequence Of the Dnmp Kinasessupporting

confidence: 71%

Section: Discussionmentioning

confidence: 99%

Section: Cloning and Over-expression Of Dnmp Kinase Genesmentioning

confidence: 98%

See 1 more Smart Citation

Cloning of deoxynucleoside monophosphate kinase genes and biosynthesis of deoxynucleoside diphosphates

Bao

Ryu²

2006

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

“…PYR6 is a uridine 5Ј-monophosphate (UMP)/cytidine 5Ј-monophosphate (CMP) kinase that converts UMP/CMPs to uridine and cytidine diphosphates, respectively. PYR6 orthologs in bacteria and yeast are required for cellular proliferation and RNA and protein synthesis (44,45). PYR6 has also been detected in the mature pollen of Arabidopsis, consistent with its role in cell proliferation and division (46).…”

Section: Fig 8 Spectral Count Analysis Of Labeled Peptidesmentioning

confidence: 64%

Profiling Protein Kinases and Other ATP Binding Proteins in Arabidopsis Using Acyl-ATP Probes

Villamor

Kaschani

Colby

et al. 2013

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Many protein activities are driven by ATP binding and hydrolysis. Here, we explore the ATP binding proteome of the model plant Arabidopsis thaliana using acyl-ATP (AcATP) 1 probes. These probes target ATP binding sites and covalently label lysine residues in the ATP binding pocket. Gel-based profiling using biotinylated AcATP showed that labeling is dependent on pH and divalent ions and can be competed by nucleotides. The vast majority of these AcATP-labeled proteins are known ATP binding proteins. Our search for labeled peptides upon in-gel digest led to the discovery that the biotin moiety of the labeled peptides is oxidized. The in-gel analysis displayed kinase domains of two receptor-like kinases (RLKs) at a lower than expected molecular weight, indicating that these RLKs lost the extracellular domain, possibly as a result of receptor shedding. Analysis of modified peptides using a gel-free platform identified 242 different labeling sites for AcATP in the Arabidopsis proteome. Examination of each individual labeling site revealed a preference of labeling in ATP binding pockets for a broad diversity of ATP binding proteins. Of these, 24 labeled peptides were from a diverse range of protein kinases, including RLKs, mitogen-activated protein kinases, and calcium-dependent kinases. A significant portion of the labeling sites could not be assigned to known nucleotide binding sites. However, the fact that labeling could be competed with ATP binding and hydrolysis are the driving processes in all living organisms. Hundreds of cellular proteins are able to bind and hydrolyze ATP to unfold proteins, transport molecules over membranes, or phosphorylate small molecules or proteins. Proteins with very different structures are able to bind ATP. A large and important class of ATP binding proteins is that of the kinases, which transfer the gamma phosphate from ATP to substrates. Kinases, and particularly protein kinases, play pivotal roles in signaling and protein regulation.The genome of the model plant Arabidopsis thaliana encodes for over 1099 protein kinases and hundreds of other ATP binding proteins (1, 2). Protein kinases are involved in nearly all signaling cascades and regulate processes ranging from cell cycle to flowering and from immunity to germination. Many protein kinases in plants are receptor-like kinases (RLKs), often carrying extracellular leucine-rich repeats (LRRs). The RLK class contains at least 610 members (3), including famous examples such as receptors involved in development (e.g. BRI1, ER, CLV1) and immunity (e.g. FLS2, EFR). Other important classes are mitogen-activated protein (MAP) kinases (MPKs) (20 different members), MPK kinase kinase kinases (MAP3Ks) (60 different members (4)), and calcium-dependent protein kinases (CPKs) (34 different members (5)). Because of their diverse and important roles, protein kinases have been intensively studied in plant science. The current approach is to study protein kinases individually-a daunting task, considering the remaining hundreds of uncharacterized p...

show abstract

“…Each enzyme catalyses the synthesis of a nucleoside diphosphate that is, in turn, converted to a nucleoside triphosphate by a non-specific nucleoside diphosphate kinase. In prokaryotes, there are five NMP kinases, one for the phosphorylation of each NMP, whereas in eukaryotic organisms, the phosphorylation of both uridine monophosphate (UMP) and cytidine monophosphate (CMP) is carried out by a bifunctional UMP/CMP kinase (Liljelund & Lacroute, 1986).…”

Section: Introductionmentioning

confidence: 99%

Structure–function relationships of UMP kinases from pyrH mutants of Gram-negative bacteria

et al. 2004

View full text Add to dashboard Cite

Bacterial uridine monophosphate (UMP) kinases are essential enzymes encoded by pyrH genes, and conditional-lethal or other pyrH mutants were analysed with respect to structure-function relationships. A set of thermosensitive pyrH mutants from Escherichia coli was generated and studied, along with already described pyrH mutants from Salmonella enterica serovar Typhimurium. It is shown that Arg-11 and Gly-232 are key residues for thermodynamic stability of the enzyme, and that Asp-201 is important for both catalysis and allosteric regulation. A comparison of the amino acid sequence of UMP kinases from several prokaryotes showed that these were conserved residues. Discussion on the enzyme activity level in relation to bacterial viability is also presented.

show abstract

Genetic characterization and isolation of theSaccharomyces cerevisiae gene coding for uridine monophosphokinase

Cited by 48 publications

References 42 publications

Cloning of deoxynucleoside monophosphate kinase genes and biosynthesis of deoxynucleoside diphosphates

Cloning of deoxynucleoside monophosphate kinase genes and biosynthesis of deoxynucleoside diphosphates

Profiling Protein Kinases and Other ATP Binding Proteins in Arabidopsis Using Acyl-ATP Probes

Structure–function relationships of UMP kinases from pyrH mutants of Gram-negative bacteria

Contact Info

Product

Resources

About