Trichinella spiralis and Trichinella pseudospiralis: developmental patterns of enzymes involved in thymidylate biosynthesis and pyrimidine salvage.
Thymidylate synthase, dihydrofolate reductase and dUTPase specific activities were found to remain at a high and constant level in crude extracts from adult worms of Trichinella spiralis, as well as from muscle larvae of both Trichinella spiralis (isolated 1-24 months after infection) and Trichinella pseudospiralis (isolated 5.5-13 months after infection). The results obtained with Trichinella pseudospiralis muscle larvae isolated with the use of pepsin did not differ from those obtained when pepsin was not used. No thymidine kinase activity could be detected in muscle larvae of either species and thymidine phosphorylase could be found only in T. pseudospiralis larvae isolated without the use of pepsin. Muscle larvae of both species contained orotidylate phosphoribosyl transferase activity, pointing to a possibility of 5-fluorouracil activation. Uridine phosphorylase, another enzyme involved in 5-fluorouracil anabolism, was also present in T. pseudospiralis muscle larvae. Results of comparative studies on inhibition of purified T. spiralis and rat thymidylate synthases by substrate (4-thio-5-fluoro-dUMP, 2-thio-5-fluoro-dCMP and N4-hydroxy-dCMP) and cofactor (ZD 9331) analogues indicated only dUMP analogues to show feeble selectivity towards the parasite enzyme. A hypothesis is discussed, assuming high expression of thymidylate synthase in muscle larvae to be connected with their cells being arrested in the cell cycle.
Rode
W
W
Nencki Institute of Experimental Biology, Polish Academy of Sciences, Warszawa, Poland. rode@nencki.gov.pl
Dabrowska
M
M
Zieliński
Z
Z
Gołos
B
B
Wranicz
M
M
Felczak
K
K
Kulikowski
T
T
D000818:Animals; D005260:Female; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007700:Kinetics; D008297:Male; D051379:Mice; D018482:Muscle, Skeletal / Q000469:parasitology*; D009962:Orotate Phosphoribosyltransferase / Q000032:analysis; D010770:Phosphotransferases / Q000032:analysis; D011743:Pyrimidines / Q000378:metabolism*; D013937:Thymidine Kinase / Q000032:analysis; D013938:Thymidine Monophosphate / Q000096:biosynthesis*; D013939:Thymidine Phosphorylase / Q000032:analysis; D013940:Thymidylate Synthase / Q000032:analysis*; D014234:Trichinella / Q000201:enzymology*; D017160:Trichinella spiralis / Q000201:enzymology*; D014543:Uridine Phosphorylase / Q000032:analysis
D011743:Pyrimidines; D013938:Thymidine Monophosphate; D013940:Thymidylate Synthase; D009962:Orotate Phosphoribosyltransferase; D014543:Uridine Phosphorylase; D013939:Thymidine Phosphorylase; D010770:Phosphotransferases; D013937:Thymidine Kinase; C021892:nucleoside phosphotransferase