Trypanosoma cruzi: a gene family encoding chitin-binding-like proteins is posttranscriptionally regulated during metacyclogenesis.
The development of the representation of differential expression method has lead to the cloning of Trypanosoma cruzi stage-specific genes. We used this method to characterize a multicopy gene family differentially expressed during metacyclogenesis. The genomic and cDNA clones sequenced encoded three short cysteine-rich polypeptides, of two types, with predicted molecular masses of 7.1, 10.4, and 10.8 kDa. We searched GenBank for similar sequences and found that the sequences of these clones were similar to that encoding the wheat germ agglutinin protein. The region of similarity corresponds to the chitin-binding domain, with eight similarly positioned half-cysteines and conserved aromatic residues involved in chitin recognition. Multiple copies of the genes of this family are present on a high- molecular-mass chromosome. We studied the expression of genes of this family during metacyclogenesis by determining messenger RNA (mRNA) levels. The mRNAs for the members of this gene family were present in the total RNA fraction but were mobilized to the polysomal fraction of adhered (differentiating) epimastigotes during metacyclogenesis, with a peak of accumulation at 24 of differentiation. Polyclonal antisera were raised against a recombinant protein and a synthetic peptide. The specific sera obtained detected 7- and 11-kDa proteins in T. cruzi total protein extracts. The 11-kDa protein was present in similar amounts in the various cell populations, whereas the 7-kDa protein displayed differential synthesis during metacyclogenesis, with maximal levels in 24-h-adhered (differentiating) epimastigotes.
Dallagiovanna
B
B
Instituto de Biologia Molecular do Paraná, IBMP, Rua Prof. Algacyr Munhoz Mader 3775, 8135-010 Curitiba, PR, Brazil.
Plazanet-Menut
C
C
Ogatta
S F
SF
Avila
A R
AR
Krieger
M A
MA
Goldenberg
S
S
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D002448:Cell Adhesion; D002686:Chitin / Q000378:metabolism; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000737:chemistry; D015870:Gene Expression; D018503:Genome, Protozoan; D007106:Immune Sera / Q000737:chemistry; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000502:physiology*; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D016053:RNA, Protozoan / Q000032:analysis; D011817:Rabbits; D014349:Trypanosoma cruzi / Q000737:chemistry / Q000235:genetics* / Q000502:physiology
D002352:Carrier Proteins; D016054:DNA, Protozoan; D007106:Immune Sera; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D002686:Chitin