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Biomedical subjects

M J Babcock

Publications and source records attributed to M J Babcock.

9 recordsLinked to original sources
PubMed1992

Regulation and secretion of an extracellular esterase from Streptomyces scabies.

Production of a heat-stable, extracellular esterase by Streptomyces scabies is regulated by zinc ions. The esterase-encoding gene (est) from S. scabies was cloned and expressed in Streptomyces lividans. In S. lividans, expression of the est gene is also regulated by Zn2+, and the esterase is efficiently secreted in this organism. The sequence of the est gene suggests that a 39-amino acid signal peptide is removed during secretion of this protein. Deletion analysis has indicated that the hydrophobic domain of the signal peptide is required for secretion. Gel retardation assays and DNaseI footprinting using an S-30 protein extract from S. scabies have previously identified a specific 23-bp protein-binding site upstream from the est coding sequence. Deletion of this protein-binding sequence significantly decreased expression of the est gene.

Amino Acid Sequence↗
PubMed1992

Molecular analysis of sporulation in Streptomyces griseus.

Previous evidence suggested that orf1590 from Streptomyces griseus has the potential to encode two polypeptide products from temporally regulated nested open frames (orfs) and that the longer polypeptide may be a DNA-binding protein. We have developed a hypothetical model of the role of orf1590 in sporulation of S. griseus and have begun to test this model by determining the nucleotide sequence of the orf1590 counterpart from Streptomyces coelicolor. The conservation of the helix-turn-helix domain and the two potential translation start codons is consistent with our model. Continued analysis of bald mutants of S. griseus has indicated that several prematurely synthesize sporulation septa and spore walls. One of these nonsporulating strains appears to be a bldA mutant of S. griseus. Complementation analysis suggests that at least three genetic loci are involved in the correct timing of deposition of sporulation septa and wall thickening.

Amino Acid Sequence↗
PubMed1992

Identification of a protein-binding sequence involved in expression of an esterase gene from Streptomyces scabies.

Expression of an esterase gene from Streptomyces scabies is regulated by zinc in both Streptomyces scabies and Streptomyces lividans. A specific protein-binding site was identified on an esterase promoter fragment by using an S-30 extract from S. scabies. The location of the protein-binding site was determined by gel shift assays of promoter deletion fragments and by DNase I footprinting analysis. The protein-binding site maps from nucleotides -59 to -81 relative to the start of transcription. An esterase gene construct cloned and expressed in S. lividans was used to assess the importance of the protein-binding site. Deletion of the 23-bp protein-binding site resulted in a 10-fold decrease in esterase production when cells were grown in zinc-inducing conditions. The protein-binding site may represent a region involved in positive regulation of the S. scabies esterase gene.

Amino Acid Sequence↗
PubMed1990

Transcriptional and translational features of a sporulation gene of Streptomyces griseus.

The nucleotide (nt) sequence of a 2.8-kb fragment of DNA that restores sporulation to one class of bald mutants of Streptomyces griseus revealed an open reading frame (ORF) with the potential to encode a 55.5-kDa polypeptide. The presence of an in-frame TTA in the coding sequence indicated that translation is likely to require the tRNA(Leu)UUA encoded by the bldA gene. Two overlapping transcripts are initiated at transcriptional start points (tsp) separated by 258 nt and are transcribed in the same direction. The downstream tsp lies within the ORF and is followed by a second potential translation initiation site, which would encode a 49.5-kDa polypeptide in the same reading frame as the 55.5-kDa polypeptide. Transcription assays suggested that both tsp functioned during vegetative growth, but the relative abundance of the shorter transcript decreased during the early stages of submerged sporulation. Analysis of sequentially deleted subclones indicated that expression of the longer ORF was necessary to complement bald mutants. The presence of two tsp alternating with potential translation start codons suggests the temporally regulated synthesis of two polypeptides that have identical C termini but different N termini.

Amino Acid Sequence↗
PubMed1988

Cloning of DNA involved in sporulation of Streptomyces griseus.

Twenty-two bald mutants of Streptomyces griseus were isolated and classified into four phenotypic groups, two of which showed conditional sporulation. A 3-kilobase fragment of DNA was cloned in a high-copy-number vector and detected by its ability to restore sporulation to one class of conditionally bald mutants. Analysis of subclones demonstrated that the sporulation property was contained within a 2.5-kilobase fragment. Hybridization studies and restriction analysis indicated that this DNA fragment was present in several Streptomyces species and was distinct from DNA that has been shown to complement afsA mutants of S. bikiniensis and bldA mutants of S. coelicolor.

Cloning, Molecular↗
PubMed1978

Hepatic transaminase in protein-restricted rats: development of a controlled model.

A marginal protein malnutrition model which makes allowances for the increasing nutritional requirements of the growing rat and the effects of dietary manipulation on diurnal rhythms, while still rigidly controlling the level of protein restriction, is reported. A predetermined, constantly increasing intake of four purified diets, providing approximately 40 to 160% of the National Research Council protein requirement for rats was fed to rats receiving a nitrogen-free energy source adlibitum. This standardization reduced within-group variation and allowed precise growth reproducibility. Biochemical parameters were measured at evenly spaced intervals throughout the day to remove diurnal differences from between-group comparisons. Growth, nitrogen balance, and 24-hour means of liver weight, DNA, RNA, protein, and glutamate-oxaloacetate and glutamate-pyruvate transaminases reflected protein intake. However, when growth was depressed 10, 30, or 60%, liver GPT/DNA was depressed 40, 49, or 67%, respectively. Hepatic GPT appears to be a sensitive and accurate indicator of marginal protein malnutrition.

Alanine Transaminase↗