Trypanosoma brucei variant-specific glycoprotein gene chromatin is sensitive to single-strand-specific endonuclease digestion.
Active variant surface glycoprotein (VSG) gene chromatin is preferentially digested by the restriction enzyme HinfI in nuclei of bloodstream variants of Trypanosoma brucei. HinfI sensitivity of VSG gene chromatin is not observed in nuclei of relapse variants in which the VSG gene has been inactivated in situ. Active VSG gene chromatin is preferentially degraded by the single-strand-specific endonucleases S1 and Bal31. This sensitivity is not the result of pre-existing single-strand breaks or a detectably altered nucleosomal organization. Trypanosome nuclei in which the run-on transcription of VSG genes has been specifically shut down have been used to show that Hinfl and Bal31 sensitivity is not dependent upon continued transcription of the VSG gene. The presence of single-stranded DNA regions within VSG gene chromatin is consistent with a model in which VSG genes are activated by increased torsional stress.
Greaves
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DR
Division of Molecular Biology, Netherlands Cancer Institute, Antoni van Leeuwenhoekhuis, Amsterdam.
Borst
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P
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics*; D002843:Chromatin / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific; D004720:Endonucleases / Q000378:metabolism*; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000235:genetics*; D009690:Nucleic Acid Conformation; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013314:Stress, Mechanical; D013379:Substrate Specificity; D014158:Transcription, Genetic; D014346:Trypanosoma brucei brucei / Q000235:genetics*
D000953:Antigens, Protozoan; D002843:Chromatin; D006023:Glycoproteins; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases; D015719:Single-Strand Specific DNA and RNA Endonucleases