TISSUE-SPECIFIC PERMUTATIONS OF SELF-CLEAVING NEWT SATELLITE-2 TRANSCRIPTS

Synthetic transcripts of satellite-2 (sat2) DNA from the newt undergo self-catalyzed, site-specific cleavage in vitro using the conserved hammerhead (HH) motif found in a number of infectious plant RNAs. We have analyzed sat2 transcripts from a variety of tissues to obtain evidence for the occurrence of self-cleavage in vivo. We found two distinct types of sat2 transcript populations. Monomeric transcripts in the testes and in somatic tissues such as the liver have ends that map to the in vitro self-cleavage site, and have end groups (5'-hydroxyls and phosphate blocked 3' ends) similar to those produced by self-cleavage. This is the first indication of HH-mediated self-cleavage occurring in animal cells. The monomeric sat2 transcripts found in the newt ovary have a different permutation of the basic DNA repeat sequence. These transcripts begin and end 47 nucleotides upstream from the in vitro self-cleavage site. Despite their apparent lack of utilization of the self-cleavage site, they have end groups which indicate that processing is involved in their formation.