Cloning of the tyrP gene and further characterization of the tyrosine-specific transport system in Escherichia coli K-12.

AUTOR(ES)

Wookey, P J

RESUMO

The tyrP gene which codes for a component of the tyrosine-specific transport system of Escherichia coli has been cloned on a 2.8-kilobase insert into plasmid pBR322. Transposon mutagenesis, using Tn1000, indicates that the tyrP+ gene is at least 1.1 kilobase in length. Labeling of the tyrP protein in maxicells with [35S]methionine indicates an apparent molecular weight of ca. 24,500. Sedimentation analysis reveals that the tyrP protein is associated with the cell membrane and is not free in the cytoplasm or periplasm. Strains with many copies of the tyrP+ gene show an enhanced uptake of tyrosine, but the expression of the system is still modulated by tyrosine and phenylalanine in the presence of the tyrR+ regulator protein. Accumulated radioactive tyrosine is rapidly effluxed by the addition either of energy uncouplers or of excess nonradioactive tyrosine, indicating that the transport system is energized by the proton motive force and that the internal pool is readily exchangeable. The effect of increasing expression of the tyrP gene on the steady-state level of tyrosine accumulated by cells indicates that although the transport system may be dependent on the proton motive force to drive uptake, the system never reaches thermodynamic equilibrium with it.

Documentos Relacionados

• DNA sequence of the gene (tyrP) encoding the tyrosine-specific transport system of Escherichia coli.
• Molecular analysis of the promoter operator region of the Escherichia coli K-12 tyrP gene.
• Characterization of the specific pyruvate transport system in Escherichia coli K-12.
• Mutational analysis of repression and activation of the tyrP gene in Escherichia coli.
• Cloning and characterization of Escherichia coli K-12 regulator gene tyrR.