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Biomedical subjects

D Patterson

Publications and source records attributed to D Patterson.

34 records · Page 2Linked to original sources
PubMed1977

Biochemical genetic analysis of pyrimidine biosynthesis in mammalian cells: I. Isolation of a mutant defective in the early steps of de novo pyrimidine synthesis.

The isolation and characterization of a new mutant of Chinese hamster ovary cells is described. This mutant, Urd-A, shows an absolute requirement for exogenously added pyrimidines for growth. Complementation analysis indicates that the lesion in this mutant is recessive. Revertants can be isolated at frequencies suggesting that it is a single gene alteration. Biochemical analysis of cell-free extracts of CHO-K1 (Urd+) and Urd-A revealed that Urd-A possesses no more than 10% of wild-type levels of carbamyl phosphate synthetase (EC 2.7.2.9) activity, no more than 1% of wild-type levels of aspartate transcarbamylase (EC 1.2.3.2) activity, and undetectable levels of dihydroorotase (EC 3.5.2.3) activity. Thus, this mutant appears simultaneously to possess marked or complete deficiencies in the activities of the first three enzymes of pyrimidine biosynthesis. Activities of the other enzymes of the pathway appear normal. The use of this mutant for biochemical-genetic studies of pyrimidine biosynthesis is discussed.

Aspartate Carbamoyltransferase↗
PubMed1976

Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism. IV. Isolation of a mutant which accumulates adenylosuccinic acid and succinylaminoimidazole carboxamide ribotide.

The production, isolation, and characterization of a new complementation group (Ade-I) of adenine-requiring mutant of Chinese hamster cells (CHO-K1) is described. This mutant accumulates two intermediates of purine biosynthesis, both of which contain an aspartate moiety. One of these is shown to be adenylosuccinic acid (AMPS) by chromatographic analysis, while evidence is presented that strongly suggests the other intermediate is succinylaminoimidazole carboxamide ribotide (SAICAR). Thus, Ade-I is most likely lacking the activity of the enzyme adenylosuccinase (EC 4.3.2.2). The use of this and similar mutants for the analysis of regulation of purine biosynthesis in mammalian cells is discussed.

Adenine↗
PubMed1976

Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism III. Isolation and characterization of a mutant unable to convert IMP to AMP.

The isolation and characterization of a new mutant of Chinese ovary cells (CHO-K1) is described. This mutant, Ade-H, has the following properties: (1) it forms a new genetic complementation group; (2) it specifically requires adenine for growth and will not grow on aminoimidazole carboxamide (AIC) or hypoxanthine; (3) it accumulates IMP; (4) it cannot synthesize adenine nucleotides; (5) its phenotype can be mimicked by treatment of CHO-K1 (the wild type parental strain) with hadacidin, an inhibitor of adenylosuccinate synthetase (E.C.6.3.4.4). Thus, the site of the defect in this mutant is presumed to involve the step in adenylate biosynthesis catalyzed by this enzyme. The usefulness of Ade-H for the study of regulation of purine biosynthesis in mammalian cells is discussed.

Adenine↗
PubMed1976

Isolation and characterization of temperature-sensitive mutants of Chinese hamster ovary cells after treatment with UV and x-irradiation.

The isolation of ten conditionally lethal temperature-sensitive mutants of the Chinese hamster ovary cell (CHO-Kl, pro-) by the BUdR-visible light selection procedure described. Treatment with radiation at doses known to cause single gene mutation in mammalian cells increases the mutation frequency by a factor of at least 14. These mutants will grow with normal plating efficiency at 34.5 degrees but will not grow at 39.5 degrees. Complementation analysis by two independent methods indicates that all mutants are recessive and allows the assignment of the mutants to six genetically independent complementation groups. Reversion analysis indicates that the TS-mutants are stable, spontaneous revertants arising at a frequency of less than 10(-6). Preliminary chromosome analysis revealed no systematic chromasomal abnormality in the mutants. Mitotic accumulation is used to study the generation time of the parental cells and representative mutants at 34.5 degrees and 39.5 degrees. The uses of these mutants for genetic analysis of mammalian cells in culture is discussed.

Cell Division↗
PubMed1976

Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism: isolation, selection, and characterization of a mutant lacking hypoxanthine-guanine phosphoribosyltransferase activity by nutritional means.

Mutants of the Chinese hamster ovary cell derived from CHO-K1 have been selected for lack of hypoxanthine-guanine phosphoribosyltransferase (EC 2.4.2.8) (HGPRT) without the use of a drug-resistance protocol. The procedure depends on the use of a parental strain carrying a mutation making it unable to synthetize purines and thus dependent upon exogenously added purines for growth. The standard "BUdR-visible-light" procedure is then used to select those cells which can use adenine but cannot use hypoxanthine as a purine source. These cells are shown to be thioguanine resistant, to be unable to incorporate exogenously added hypoxanthine into purine nucleotides, to complement our other adenine-specific purine auxotrophs, Ade-H and Ade-I but not to complement a cell isolated by virtue of thioguanine resistance, and to lack the activity of HGPRT. The use of such multiply marked mutants and cells related to them for further analysis of purine nucleotide biosynthesis and interconversion is discussed.

Animals↗
PubMed1975

Biochemical genetics of Chinese hamster cell mutants with deviant purine metabolism: biochemical analysis of eight mutants.

Purine biosynthesis was studied in 8 mutants of Chinese hamster cells which require purines for growth and in wild-type cells which do not show this nutritional requirement. Of these, 6 mutants, ade-B, ade-D, ade-E, ade-F, GAT-, and AT-, were shown to accumulate metabolic intermediates not accumulated by wild-type cells. These intermediates were shown to be compounds unique to the adenylic acid biosynthetic pathway by the following criteria: (a) their radioisotopic labeling properties, (b) their response to agents which specifically inhibit known enzymatic steps in the pathway, (c) their chromatographic properties, and (d) spectrophotometric analysis. Two mutants, ade-A and ade-C, accumulate no detectable compounds not accumulated by the wild type. These 2 mutants are believed to be defective in steps very early in the purine biosynthetic pathway. The sites of the defects in the other mutants are proposed, and the usefulness of these mutants is discussed.

Adenosine Monophosphate↗