TY - JOUR
T1 - Sea urchin sperm chromatin structure as probed by pancreatic DNase I
T2 - Evidence for a novel cutting periodicity
AU - Arceci, Robert J.
AU - Gross, Paul R.
N1 - Funding Information:
We would like to thank Dr. Martin Gorovsky, Dr. Robert Angerer, and Mr. Stuart Moss for continuing and helpful discussions of this work. The computer program used in analyzing the nuclease kinetic data was kindly made available by Dr. Robert Angerer. We are grateful to Dr. Claiborne Glover, who carried out the reconstitutions and prepared the grids used for electron microscopy. Support has been provided by grants from the National Institute of Child Health and Human Development (HD-08652) and the Rockefeller Foundation (GA PD 7812).
PY - 1980
Y1 - 1980
N2 - Chromatin from mature sea urchin spermatozoa is highly compacted and composed almost entirely of DNA and the five histones, although sperm H1, H2A, and H2b histones differ from those found in embryo or somatic cell nuclei. Release of acid-soluble DNA during pancreatic DNase I digestion is 20-fold slower from sperm nuclei than from embryonic nuclei. Following DNase I digestion, most sperm nuclear DNA remains at high molecular weight, although there appears to be some release of 10 base oligomer fragments. Size analysis of the higher molecular weight DNA reveals a series of fragments that indicate a cutting periodicity of approximately 500 base pairs. This pattern remains when electrophoretic separation is carried out under denaturing conditions. The 500 base pair cleavage pattern was not detected in digestions of embryonic nuclei. Nucleosomes reconstituted with fractionated core histones from sperm gave, upon digestion, a characteristic 10 base “ladder,” with no resistant high molecular weight DNA. Micrococcal nuclease and DNase II digested sperm nuclei to produce DNA fragments with a calculated repeat length of 248 ± 3 and 246 ± 6 base pairs, respectively. The structural basis for the 500 base pair cutting periodicity in sperm nuclei may reside in the unique sperm H1 histone.
AB - Chromatin from mature sea urchin spermatozoa is highly compacted and composed almost entirely of DNA and the five histones, although sperm H1, H2A, and H2b histones differ from those found in embryo or somatic cell nuclei. Release of acid-soluble DNA during pancreatic DNase I digestion is 20-fold slower from sperm nuclei than from embryonic nuclei. Following DNase I digestion, most sperm nuclear DNA remains at high molecular weight, although there appears to be some release of 10 base oligomer fragments. Size analysis of the higher molecular weight DNA reveals a series of fragments that indicate a cutting periodicity of approximately 500 base pairs. This pattern remains when electrophoretic separation is carried out under denaturing conditions. The 500 base pair cleavage pattern was not detected in digestions of embryonic nuclei. Nucleosomes reconstituted with fractionated core histones from sperm gave, upon digestion, a characteristic 10 base “ladder,” with no resistant high molecular weight DNA. Micrococcal nuclease and DNase II digested sperm nuclei to produce DNA fragments with a calculated repeat length of 248 ± 3 and 246 ± 6 base pairs, respectively. The structural basis for the 500 base pair cutting periodicity in sperm nuclei may reside in the unique sperm H1 histone.
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U2 - 10.1016/0012-1606(80)90509-6
DO - 10.1016/0012-1606(80)90509-6
M3 - Article
C2 - 7439531
AN - SCOPUS:0019087066
SN - 0012-1606
VL - 80
SP - 210
EP - 224
JO - Developmental biology
JF - Developmental biology
IS - 1
ER -