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Nucleic Acids Research, 1994, Vol. 22, No. 6 1029-1036
© 1994

MOLECULAR BIOLOGY

A group III twintron encoding a maturase-like gene excises through lariat intermediates

Donald W. Copertino1,2,+, Edina T. Hall2,§, Fredrick W. Van Hook2, Kristin P. Jenkins2 and Richard B. Hallick1,2,*

1Departments of Biochemistry, University of Arizona Tucson, AZ 85721, USA 2Molecular and Cellular Biology, University of Arizona Tucson, AZ 85721, USA

* To whom correspondence should be addressed

Received December 9, 1993. Revised February 10, 1994. Accepted February 10, 1994.

The 1605 bp intron 4 of the Euglena gracills chloroplast psbC gene was characterized as a group III twintron composed of an internal 1503 nt group III intron with an open reading frame of 1374 nt [ycf13, 458 amino acids), and an external group III intron of 102 nt. Twintron excision proceeds by a sequential splicing pathway. The splicing of the internal and external group III introns occurs via lariat intermediates. Branch sites were mapped by primer extension RNA sequencing. The unpaired adenosines in domains VI of the internal and external introns are covalently linked to the 5' nucleotlde of the intron via 2' -5' phosphodlester bonds. This bond is susceptible to hydrolysis by the debranchlng activity of the HeLa nuclear S100 fraction. The internal intron and presumptive ycf13 mRNA accumulates primarily as a linear RNA, although a lariat precursor can also be detected. The ycf13 13 gene encodes a maturase-like protein that may be involved In group III Intron metabolism.

Present addresses: + Department of Neurobiology, The Scnpps Research Institute, 10666 North Torrey Pines Road, La Jolla, CA 92037

Present addresses: §Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309, USA


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