Bending of DNA segments with Saccharomyces cerevisiae autonomously replicating sequence activity, isolated from basidiomycete mitochondrial linear plasmids |
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Authors: | Manahu Nakajima Qaiser I. Sheikh Kazuyoshi Yamaoka Yoshiyasu Yui Susumu Kajiwara Kazuo Shishido |
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Affiliation: | (1) Department of Life Science, Tokyo Institute of Technology, Nagatsuta, Midori-ku, 227 Yokohama, Japan;(2) Present address: Department of Chemistry, Islamia University, Bahawalpur, Pakistan;(3) Present address: Biotechnology Research Laboratory, Teijin Limited, Asahigaoka, Hino-shi, 191 Tokyo, Japan;(4) Present address: First Research Center, Technical Research and Development Institute, Nakameguro, Meguro-ku, 153 Tokyo, Japan Defense Agency, Japan |
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Abstract: | Summary Previous studies have indicated that DNA bending is a general structural feature of sequences (ARSs) from cellular DNAs of yeasts and nuclear and mitochondrial genomic DNAs of other eukaryotes that are capable of autonomous replication in Saccharomyces cerevisiae. Here we showed that bending activity is also tightly associated with S. cerevisiae ARS function of segments cloned from mitochondrial linear DNA plasmids of the basidiomycetes Pleurotus ostreatus and Lentinus edodes. Two plasmids, designated pLPO2-like (9.4 kb), and pLPO3 (6.6 kb) were isolated from a strain of P. ostreatus. A 1029 by fragment with high-level ARS activity was cloned from pLPO3 and it contained one ARS consensus sequence (A/T)TTTAT(A/G)TTT(A/T) indispensable for activity and seven dispersed ARS consensus-like (10/11 match) sequences. A discrete bent DNA region was found to lie around 500 by upstream from the ARS consensus sequence (T-rich strand). Removal of the bent DNA region impaired ARS function. DNA bending was also implicated in the ARS function associated with a 1430 by fragment containing three consecutive ARS consensus sequences which had been cloned from the L. edodes plasmid pLLE1 (11.0 kb): the three consecutive ARSs responsible for high-level ARS function occurred in, and immediately adjacent to, a bent DNA region. A clear difference exists between the two plasmid-derived ARS fragments with respect to the distance between the bent DNA region and the ARS consensus sequence(s). |
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Keywords: | DNA bending Autonomously replicating sequence Saccharomyces cerevisiae Basidiomycete Linear plasmid |
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