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Infection and Immunity, November 1999, p. 6157-6160, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Analysis of Pneumocystis carinii Introns

Charles F. Thomas Jr.,1,* Edward B. Leof,1,2 and Andrew H. Limper1,2

Thoracic Diseases Research Unit, Division of Pulmonary, Critical Care and Internal Medicine, Department of Medicine,1 and Department of Biochemistry and Molecular Biology,2 Mayo Clinic and Foundation, Rochester, Minnesota 55905

Received 29 March 1999/Returned for modification 17 June 1999/Accepted 10 August 1999

Pneumocystis carinii is an ascomycete phylogenetically related to Schizosaccharomyces pombe. Little is known about gene regulation in P. carinii. The removal of introns from pre-mRNA requires spliceosomal recognition of the intron-exon boundary. In S. pombe and higher eukaryotes, this boundary and a branch site within the intron are conserved. We recently demonstrated that P. carinii cdc2 cDNA can complement S. pombe containing conditional mutations of cdc2, an essential gene involved in cell cycle regulation. We next tested whether P. carinii genomic cdc2 (with six introns) could also complement S. pombe cdc2 mutants and found genomic sequences incapable of this activity. Reverse transcriptase PCR confirmed the inability of the S. pombe cdc2 mutants to splice the P. carinii genomic cdc2. Analysis of 83 introns from 19 P. carinii protein-encoding genes demonstrated that the sequence GTWWDW functions as a donor consensus in P. carinii, whereas YAG serves as an acceptor consensus. These sequences are similar in S. pombe; however, a branch site sequence was not found in the P. carinii genes studied.

* Corresponding author. Mailing address: Thoracic Diseases Research Unit, 696 Guggenheim Building, Mayo Clinic and Foundation, Rochester, MN 55905. Phone: (507) 284-2301. Fax: (507) 284-4521. E-mail: thomas.charles{at}mayo.edu.

Infection and Immunity, November 1999, p. 6157-6160, Vol. 67, No. 11
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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