Studies of elongation factor Tu in Streptococcus faecium (ATCC 9790)

Thesis/Dissertation · Wed Jan 01 00:00:00 EST 1986

OSTI ID:6619215

Bourbeau, P P

It has been known for over twenty years that elongation factor Tu (Ef-Tu) is one of the proteins involved in protein synthesis in bacteria. Several years ago, it was proposed that Ef-Tu may, in addition, have other structural functions in bacterial. The author's research has examined the function of Ef-Tu in Streptococcus faecium. Using an antibiotic kirromycin, which specifically inhibits Ef-Tu function, the effects upon a number of cellular parameters were determined. Inhibition of both protein and RNA synthesis was found to be similar to the effect of chloramphenicol. Using the residual division technique for the determination of cell cycle events with both heterogeneous and sucrose gradient fractionated cell populations, a kirromycin sensitive event was detected between 8 min. (Td = 30 min.) and 19 min. (Td = 175 min.) later in the cell cycle than the chloramphenical sensitive event. This suggests that kirromycin is inhibiting a terminal cell cycle event which is in addition to the inhibition of protein synthesis. Purification of Ef-Tu was performed using two different methods: ion exchange and molecular exclusion chromatography; and GDP affinity chromatography. Various schemes were employed to try and obtain optimum cellular fractionation, allowing for both proper separation of ribosomes from the other cellular fractions and retention of enzymatic activity by Ef-Tu as determined by a /sup 3/H-GDP binding assay. Analysis of the cell cycle of S. faecium using the residual division technique was also performed. In addition, certain cell wall antibiotics were used to determine if other cell cycle events could be determined using the residual division technique.

• Research Organization: Temple Univ., Philadelphia, PA (USA)
• OSTI ID: 6619215
• Resource Relation: Other Information: Thesis (Ph. D.)
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
BIOSYNTHESIS
STREPTOCOCCUS
BIOCHEMISTRY
TRITIUM COMPOUNDS
ANTIBIOTICS
CELL CYCLE
GUANYLIC ACID
ION EXCHANGE CHROMATOGRAPHY
RNA
ANTI-INFECTIVE AGENTS
BACTERIA
CHEMISTRY
CHROMATOGRAPHY
DRUGS
LABELLED COMPOUNDS
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOTIDES
ORGANIC COMPOUNDS
SEPARATION PROCESSES
SYNTHESIS
550201* - Biochemistry- Tracer Techniques