Worm Breeder's Gazette 14(4): 40 (October 1, 1996)

These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.

Cuticlin genes organization in Ascaris lumbricoides.

Mohammed Timinouni, Claudia Paiva Nunes, Paolo Bazzicalupo

International Institute of Genetics and Biophysics, via G . Marconi 10, 80125 Napoli, Italy - bazzicalupo@iigbna.iigb.na.cnr.it

The cuticle is a central element in the structural organization of all
nematodes. It is a multi-layered structure functioning as an exoskeleton
and protecting the animal from the environment under a variety of
conditions. The cuticle is composed of collegen and an insoluble residues
named cuticlins.
        The first two cuticlin genes (cut-1 and cut-2) have been
molecularly identified and cloned from Caenorhabditis elegans.
           We have identified and cloned three cut-1  homologues from the
intestinal parasite Ascaris lumbricoides .
           Partial DNA sequencing revealed that all three genes contain at
least a domain corresponding to the middle of exon three of cut-1  of C .
           Two of the genes are separated by about 10 Kb and have opposite
directions. The other gene comes from a different genomic region or at
least from one that is more than 15 Kb away. This latter gene was studied
in more details.
           Using a RT-PCR and RACE protocols we determined the structure
the messenger which is 1687 nucleotides long and is transpliced to SL1.
Comparison of 5800 bp of genomic sequence with those of RACE clones shows
that the gene contains four exons of 188, 390, 467, and 582 bp. The
of the first, second and third introns are 1906, 378, and 413 bp
respectively. The gene codes for a predicted protein of 386 amino-acids.
           In the RNA from intestine, which is the tissue from which RACE
clones were generated, unexpectedly we have found shorter mRNAs with SL1
spliced onto the second and third exons. These shorter mRNAs are almost as
abundant as the long one. At present we do not know the functional
relevance of these mRNAs
            CUT-1 of Ascaris lumbricoides , C . elegans  and Meloidogyne
artiella  are respectively 87% and 84% identical at the amino - acids
            Like in C . elegans  and in Meloidogyne artiella  CUT-1 of
Ascaris lumbricoides  has a signal peptide. The number and the positions
all cysteine residues are conserved in the three proteins.
            We have raised antisera in rabbits against a portion of the
protein  produced in E. coli and have affinity purified specific
antibodies. We are using these in western blot and immuno-microscopy
experiments to determine the stages and tissues where the protein is