Digestion of the chicken beta-globin gene chromatin with micrococcal nuclease reveals the presence of an altered nucleosomal array characterized by an atypical ladder of DNA fragments

EMBO J. 1986 Feb;5(2):293-300.

Abstract

The structure of the chicken adult beta-globin gene chromatin in immature and mature erythrocyte nuclei has been analysed using micrococcal nuclease digestion. The resulting DNA fragments were blotted onto DBM-papers and probed with labelled DNA fragments spanning the adult beta-globin gene and its 5'- and 3'-flanking regions. The structure of the nucleosomes within and in the regions flanking the adult beta-globin gene appears to be altered in at least two ways in erythrocyte chromatin, when compared with either bulk or inactive ovalbumin gene chromatin. First, oligomeric DNA fragments containing the beta-globin gene are released faster than those of either bulk or ovalbumin gene chromatin. Second, although the difference in size of the liberated oligomeric DNA fragments is similar to the nucleosomal repeat length of bulk and ovalbumin gene chromatin, the individual oligomers are approximately 100 bp shorter than their bulk or ovalbumin gene counterparts, most noticeably when the nuclease digestion is performed at 37 degrees C. This results in an atypical ladder of approximately 300, 500, 700, 900 bp instead of the canonical chicken erythrocyte ladder which is an integral multiple of 207 bp. The same ladder was obtained from immature erythrocytes, in which the beta-globin gene is actively transcribed, and from mature erythrocytes, in which it is considered to be inactive with RNA polymerase molecules clustered in the 5' moiety of the gene. This indicates that the alteration of the nucleosomal structure is not due to transcription per se.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chickens
  • Chromatin / ultrastructure*
  • DNA / isolation & purification*
  • DNA Restriction Enzymes
  • Erythrocytes / metabolism
  • Female
  • Genes*
  • Globins / genetics*
  • Micrococcal Nuclease / metabolism*
  • Nucleosomes / ultrastructure*
  • Oligodeoxyribonucleotides / analysis
  • Oviducts / metabolism
  • Reticulocytes / metabolism
  • Spermidine / pharmacology
  • Spermine / pharmacology

Substances

  • Chromatin
  • Nucleosomes
  • Oligodeoxyribonucleotides
  • Spermine
  • Globins
  • DNA
  • DNA Restriction Enzymes
  • Micrococcal Nuclease
  • Spermidine