A. Mirandavizuete et al., THE LEVELS OF RIBONUCLEOTIDE REDUCTASE, THIOREDOXIN, GLUTAREDOXIN-1, AND GSH ARE BALANCED IN ESCHERICHIA-COLI K12, The Journal of biological chemistry, 271(32), 1996, pp. 19099-19103
The dithiol forms of thioredoxin and glutaredoxin are hydrogen donors
for ribonucleotide reductase, We have determined the intracellular lev
els of ribonucleotide reductase (RRase), thioredoxin (Trx), glutaredox
in 1 (Grx1), and glutathione (GSH) and the glutathione redox status in
new Escherichia coli K12 strains lacking thioredoxin (trxA(-)), gluta
redoxin 1 (grxA(-)), and/or GSH (gshA(-)) or overproducing Trx or Grx1
from multicopy plasmids, We propose a regulatory network in which RRa
se levels are balanced with those of Trx, Grx1, and GSH so that defici
ency or overproduction of one component would promote the opposite eff
ect on the others to maintain a balanced supply of deoxyribonucleotide
s. GSH deficiency strongly increased both Grx1 levels and RRase activi
ty, even more than Trx deficiency, Double gshA(-)trxA(-) bacteria were
viable, whereas additional deficiency in lipoate synthesis (gshA(-)tr
xA(-)lipA(-)) caused the inability to grow in minimal medium plates su
pplemented with acetate plus succinate instead of lipoic acid, Thus, l
ipoate might be the only substitute of GSH for glutaredoxin reduction
in gshA(-)trxA(-) cells, although the extremely high Grx1 content (55-
fold) of these bacteria suggests that electron transfer from lipoate m
ight be an inefficient reduction mechanism of glutaredoxins. Moreover,
the enhanced Grx1 level of gshA(-)trxA(-) cells could obviate the nee
d for a large increase in RRase activity, in contrast to grxA(-)trxA(-
) double mutant cells, Impairment of the sulfate assimilation pathway,
leading to very low GSH concentrations, and an oxidized glutathione r
edox state might explain the inability of grxA(-)trxA(-) cells to grow
in minimal medium, Restoration of nearly normal levels of both GSH co
ntent and redox status cure the growth defect.