Bands, interbands and puffs in native Drosophila polytene chromosomes are recognized by a monoclonal antibody to an epitope in the carboxy-terminal tail of histone H1.
A monoclonal antibody was raised against Drosophila melanogaster histone H1. Immunoscreening of proteolytic cleavage fragments of H1 and of a set of all possible overlapping synthetic octapeptides corresponding to the amino acid sequence of H1, revealed that the antibody recognizes an epitope within the sequence 207VTAAKPKA214 near the centre of the carboxy-terminal tail. This antibody gives positive immunofluorescence over the entire length of native D. melanogaster polytene chromosomes isolated from salivary glands by microdissection at physiological pH and ionic strength. Bands, interbands and puffs are all seen to contain H1. The immunofluorescence over puffs, albeit lower than that over bands and interbands, indicates that chromatin decondensation can occur without complete loss of H1 in these structures. The reaction of the antibody with bands suggests that the segment of the C-terminal tail containing the epitope may be exposed in the condensed 30 nm chromatin filament.
Hill
R J
RJ
CSIRO Division of Biotechnology, Laboratory for Molecular Biology, North Ryde, NSW, Australia.
Watt
F
F
Wilson
C M
CM
Fifis
T
T
Underwood
P A
PA
Tribbick
G
G
Geysen
H M
HM
Thomas
J O
JO
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002843:Chromatin / Q000032:analysis*; D002875:Chromosomes / Q000032:analysis*; D004331:Drosophila melanogaster / Q000235:genetics*; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D006657:Histones / Q000032:analysis* / Q000276:immunology; D015151:Immunoblotting; D007399:Interphase; D008969:Molecular Sequence Data; D012469:Salivary Glands; D013728:Terminator Regions, Genetic
D000911:Antibodies, Monoclonal; D002843:Chromatin; D000939:Epitopes; D006657:Histones