Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae

P. O. Weislogel; J. H.J. Hoeijmakers; A. H. Fairlamb; C. M. Kleisen; P. Borst

doi:10.1016/0005-2787(77)90180-0

Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae

P. O. Weislogel, J. H.J. Hoeijmakers, A. H. Fairlamb, C. M. Kleisen, P. Borst

Onderzoeksoutput: Bijdrage aan tijdschrift › Artikel › peer review

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1. We have used the restriction endonucleases EcoRI and PstI to further characterize the structural components of intact kinetoplast DNA networks from stationary phase Crithidia luciliae. 2. Endonuclease PstI cuts less than 7% of the mini-circles (the major component in the network) and appears to give a single cut in the maxi-circles, allowing the isolation of this minor component in linearized form. The molecular weight of these linearized maxi-circles, determined by electron microscopy with phage PM2 DNA as internal standard, is 22 · 10⁶. 3. Electron micrographs of intact networks show DNA considerably longer than the mini-circle contour length (0.8 μm) either in the network or attached to the edge. This long DNA never exceeds the size of maxi-circles (10.2 μm) and it is completely removed by treatment with either endonuclease PstI or endonuclease EcoRI (which cuts the maxi-circles and about 20% of the mini-circles). We conclude that the long DNA in these networks represent maxi-circles and that long circular oligomers of mini-circles are (virtually) absent. 4. Complete removal of maxi-circles with endonucleases PstI or EcoRI has little effect on the overall structure of the network. This shows that the mini-circles are not held together on maxi-circle strings.

Originele taal-2	Engels
Pagina's (van-tot)	167-179
Aantal pagina's	13
Tijdschrift	BBA Section Nucleic Acids And Protein Synthesis
Volume	478
Nummer van het tijdschrift	2
DOI's	https://doi.org/10.1016/0005-2787(77)90180-0
Status	Gepubliceerd - 20 sep. 1977
Extern gepubliceerd	Ja

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10.1016/0005-2787(77)90180-0

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@article{942132d0c97b4643939de25f76cfc865,

title = "Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae",

abstract = "1. We have used the restriction endonucleases EcoRI and PstI to further characterize the structural components of intact kinetoplast DNA networks from stationary phase Crithidia luciliae. 2. Endonuclease PstI cuts less than 7% of the mini-circles (the major component in the network) and appears to give a single cut in the maxi-circles, allowing the isolation of this minor component in linearized form. The molecular weight of these linearized maxi-circles, determined by electron microscopy with phage PM2 DNA as internal standard, is 22 · 106. 3. Electron micrographs of intact networks show DNA considerably longer than the mini-circle contour length (0.8 μm) either in the network or attached to the edge. This long DNA never exceeds the size of maxi-circles (10.2 μm) and it is completely removed by treatment with either endonuclease PstI or endonuclease EcoRI (which cuts the maxi-circles and about 20% of the mini-circles). We conclude that the long DNA in these networks represent maxi-circles and that long circular oligomers of mini-circles are (virtually) absent. 4. Complete removal of maxi-circles with endonucleases PstI or EcoRI has little effect on the overall structure of the network. This shows that the mini-circles are not held together on maxi-circle strings.",

author = "Weislogel, {P. O.} and Hoeijmakers, {J. H.J.} and Fairlamb, {A. H.} and Kleisen, {C. M.} and P. Borst",

note = "Funding Information: We thank Mrs. F. Fase-Fowler, Mrs. N. Van Harten-Loosbroek, Dr. J.L. Talen and Dr. R.A. Flavell for providing us with restriction endonucleases. We are indebted to the staff of the Laboratory of Electron Microscopy of the University of Amsterdam for their generous provision of facilities and to Mrs. J. Rapha~l-Snijer, Miss M.A. De Jong, Dr. M. Meijer, Dr. N. Nanninga and Dr. P.J. Weijers for help and advice in the electron microscopical work. P.O.W. was supported by an EMBO Long-Term Fellowship (1974--1975) and a post-doctoral fellowship from the National Institutes of Health, U.S.A. (GM 05338-01) (1975--1976). A.H.F. was supported by a Medical Research Council (U.K.) Travelling Fellowship.",

year = "1977",

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doi = "10.1016/0005-2787(77)90180-0",

language = "English",

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T1 - Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae

AU - Weislogel, P. O.

AU - Hoeijmakers, J. H.J.

AU - Fairlamb, A. H.

AU - Kleisen, C. M.

AU - Borst, P.

N1 - Funding Information: We thank Mrs. F. Fase-Fowler, Mrs. N. Van Harten-Loosbroek, Dr. J.L. Talen and Dr. R.A. Flavell for providing us with restriction endonucleases. We are indebted to the staff of the Laboratory of Electron Microscopy of the University of Amsterdam for their generous provision of facilities and to Mrs. J. Rapha~l-Snijer, Miss M.A. De Jong, Dr. M. Meijer, Dr. N. Nanninga and Dr. P.J. Weijers for help and advice in the electron microscopical work. P.O.W. was supported by an EMBO Long-Term Fellowship (1974--1975) and a post-doctoral fellowship from the National Institutes of Health, U.S.A. (GM 05338-01) (1975--1976). A.H.F. was supported by a Medical Research Council (U.K.) Travelling Fellowship.

PY - 1977/9/20

Y1 - 1977/9/20

N2 - 1. We have used the restriction endonucleases EcoRI and PstI to further characterize the structural components of intact kinetoplast DNA networks from stationary phase Crithidia luciliae. 2. Endonuclease PstI cuts less than 7% of the mini-circles (the major component in the network) and appears to give a single cut in the maxi-circles, allowing the isolation of this minor component in linearized form. The molecular weight of these linearized maxi-circles, determined by electron microscopy with phage PM2 DNA as internal standard, is 22 · 106. 3. Electron micrographs of intact networks show DNA considerably longer than the mini-circle contour length (0.8 μm) either in the network or attached to the edge. This long DNA never exceeds the size of maxi-circles (10.2 μm) and it is completely removed by treatment with either endonuclease PstI or endonuclease EcoRI (which cuts the maxi-circles and about 20% of the mini-circles). We conclude that the long DNA in these networks represent maxi-circles and that long circular oligomers of mini-circles are (virtually) absent. 4. Complete removal of maxi-circles with endonucleases PstI or EcoRI has little effect on the overall structure of the network. This shows that the mini-circles are not held together on maxi-circle strings.

AB - 1. We have used the restriction endonucleases EcoRI and PstI to further characterize the structural components of intact kinetoplast DNA networks from stationary phase Crithidia luciliae. 2. Endonuclease PstI cuts less than 7% of the mini-circles (the major component in the network) and appears to give a single cut in the maxi-circles, allowing the isolation of this minor component in linearized form. The molecular weight of these linearized maxi-circles, determined by electron microscopy with phage PM2 DNA as internal standard, is 22 · 106. 3. Electron micrographs of intact networks show DNA considerably longer than the mini-circle contour length (0.8 μm) either in the network or attached to the edge. This long DNA never exceeds the size of maxi-circles (10.2 μm) and it is completely removed by treatment with either endonuclease PstI or endonuclease EcoRI (which cuts the maxi-circles and about 20% of the mini-circles). We conclude that the long DNA in these networks represent maxi-circles and that long circular oligomers of mini-circles are (virtually) absent. 4. Complete removal of maxi-circles with endonucleases PstI or EcoRI has little effect on the overall structure of the network. This shows that the mini-circles are not held together on maxi-circle strings.

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DO - 10.1016/0005-2787(77)90180-0

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Characterization of kinetoplast DNA networks from the insect trypanosome Crithidia luciliae

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