Regulation of hsp70 expression in Trypanosoma cruzi by temperature and growth phase.
The steady-state level of the hsp70 mRNAs of Trypanosoma cruzi cultured at different temperatures and growth conditions has been analyzed by Northern blotting. We show that only one size class of hsp70 mRNA, of about 2.2 kb, is transcribed from the hsp70 cluster and that its transcription is constitutive at 28 degrees C. However, after a heat shock treatment at 37 degrees C for 2 h of logarithmically growing parasites, the abundance of the hsp70 mRNA increased about 4-fold. A similar increase was observed at 28 degrees C when the parasite culture reached the stationary phase of growth. On the other hand, a heat shock at 42 degrees C did not change the steady state level of the 2.2-kb size class of hsp70 mRNA. However, accumulation of transcripts of high molecular weight was detected when stationary growing parasites were cultured at 42 degrees C for 2 h. Also at 37 degrees C the steady state level of the alpha- and beta-tubulin mRNAs of logarithmically growing parasites exhibited a slight increase but only after a period of 24 h. Analysis by one-dimensional immunoblots of the Hsp70 levels showed that at 37 degrees C the abundance of the protein was 4-fold higher than at 28 degrees C. Immunoblots of high-resolution two-dimensional gel electrophoresis showed, moreover, that various isoforms of this protein are constitutively expressed at 28 degrees C and that some of them have a specific pattern of induction at 37 degrees C. We observed, moreover, that the heat shock induces the expression of a series of proteins while it causes repression of others.
Requena
J M
JM
Centro de Biología Molecular (CSIC), Universidad Autónoma de Madrid, Spain.
Jimenez-Ruiz
A
A
Soto
M
M
Assiego
R
R
Santarén
J F
JF
Lopez
M C
MC
Patarroyo
M E
ME
Alonso
C
C
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014349:Trypanosoma cruzi / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014404:Tubulin / Q000235:genetics
D006360:Heat-Shock Proteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D016053:RNA, Protozoan; D014404:Tubulin