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Nucleic Acids Research, 1991, Vol. 19, No. 23 6441-6447
© 1991

MOLECULAR BIOLOGY

The Trypanosoma brucei DNA polymerase {alpha} core subunit gene is developmentally regulated and linked to a constitutively expressed open reading frame

Peter A.J. Leegwater1,2,3,*, Marijke Strating1,2, Noel B. Murphy3, R. Frank Kooy1, Peter C. van der Vliet2 and J. Prosper Overdulve1

1Institute of Infectious Diseases and Immunology, Department of Tropical Veterinary Medicine and Protozoolgy, University of Utrecht PO Box 80165, 3508 TD Utrecht 2Laboratory for Physiological Chemistry, University of Utrecht Vondellaan 24A, 3521 GG Utrecht, The Netherlands 3International Laboratory for Research on Animal Disease PO Box 30709, Nairobi, Kenya

*To whom correspondence should be addressed at: EC Slater Institute, Department of Biochemistry, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands

Received September 6, 1991. Accepted November 13, 1991.

As an initial step towards the characterization of replicative DNA polymerases of trypanosomes, we have cloned, sequenced and examined the expression of the Trypanosoma (Trypanozoon) brucei brucei gene that encodes the DNA polymerase {alpha} catalytic core (pol{alpha}). The protein sequence contains the six conserved regions that have been recognized previously in eukaryotic and viral replicative DNA polymerases. In addition, we have identified a seventh region which appears to be conserved primarily in {alpha}-type DNA polymerases. The T.brucei DNA pol {alpha} core N-terminus is 123 and 129 amino acids smaller than that of the human and yeast homologue, respectively. The gene is separated by 386 bp from an upstream open reading frame (ORF) of 442 codons. Stable transcripts of the upstream sequence are detected in both dividing and non-dividing forms, while pol{alpha} transcripts are detected principally in dividing forms. Allelic copies of the T.brucei pol{alpha} region exhibit restriction site polymorphisms; one such sequence polymorphism affects the amino acid sequence of the T.brucei DNA pol{alpha} core. The T.brucei pol{alpha} region cross-hybridizes weakly with that of T.(Nannomonas) congolense and T.(Duttonella) vivax.


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