Crithidia luciliae: factors affecting the expression of 3'-nucleotidase/nuclease activity.
Crithidia luciliae, a trypanosomatid protozoan readily grown in axenic cultures, was shown to possess low levels of a surface membrane-bound ectoenzyme capable of hydrolyzing both 3'-ribonucleotides and nucleic acids. The specific activities of this 3'-nucleotidase/nuclease, with both mononucleotide and nucleic acid substrates, were greatly enhanced when the protozoa were deprived of purines, an essential nutrient. The catalytic activities were exhibited by a polypeptide which migrated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an Mr of 47,000. Starvation of these cells for inorganic phosphate (Pi), in media with or without purines, also led to an increase in the specific activity of the ectoenzyme compared to that of Pi- and purine-replete cells. In contrast, the level of enzyme activity was not increased when the protozoa were starved, under purine-replete conditions, for either arginine or hemin, two other essential nutrients. Cells starved simultaneously for either of the latter two nutrients and for purines also did not show increased levels of the 3'-nucleotidase/nuclease. The activation of the enzyme was also prevented by sodium arsenite, cycloheximide, actinomycin D, and tunicamycin indicating that the activation presumably required metabolic energy as well as new transcription, translation, and protein modification. The results demonstrate that the control of 3'-nucleotidase/nuclease expression is a regulated, adaptive response to growth-limiting levels of essential nutrients.
Gottlieb
M
M
Department of Immunology and Infectious Diseases, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
Mackow
M C
MC
Neubert
T A
TA
D000241:Adenosine / Q000378:metabolism; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001151:Arsenic / Q000494:pharmacology; D018053:Arsenites; D003421:Crithidia / Q000201:enzymology*; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D006427:Hemin / Q000378:metabolism; D006918:Hydroxyurea / Q000494:pharmacology; D008970:Molecular Weight; D009708:Nucleotidases / Q000378:metabolism*; D011687:Purines / Q000378:metabolism; D017670:Sodium Compounds; D013379:Substrate Specificity; D014415:Tunicamycin / Q000494:pharmacology
D018053:Arsenites; D003470:Culture Media; D011687:Purines; D017670:Sodium Compounds; D014415:Tunicamycin; D003609:Dactinomycin; C017947:sodium arsenite; D006427:Hemin; D001120:Arginine; D003513:Cycloheximide; D009708:Nucleotidases; C040470:3'-nucleotidase; D000241:Adenosine; D001151:Arsenic; D006918:Hydroxyurea