Two AT-rich satellite DNAs in the chironomid Glyptotendipes barbipes (Staeger): isolation and localization in polytene chromosomes of G. barbipes and Chironomus thummi

Abstract

Two AT-rich satellite DNAs are present in the genome of Glyptotendipes barbipes. The two satellites have densities of 1,680 g/cm3 (=21% GC) and of 1.673 g/cm3 (=13% GC) in neutral CsCl-density gradients. The main band DNA has a density of 1.691 g/cm# (= 32% GC). This value is in agreement with the 33% GC=content of G. barbipes DNA calculated from thermal denaturation (TM=83 degrees C). - In brain DNA as well as in salivary gland DNA the two satellite sequences together comprise 12-15% of the total G. barbipes DNA. Comparisons of the density profiles of DNA extracted from polytene and non-polytene larval tissue gave no hints for under-replication of the satellite DNAs during polytenization. - The two satellite DNAs have been isolated from total DNA by Hoechst 33258-CsCl density centrifugation and then localized in the polytene salivary gland chromosomes by in situ hybridization. Both satellite sequences hybridize to all heterochromatic centromer bands of all four chromosomes of G. barbipes. Satellite I (1.673 g/cm3) hybridizes mainly with the middle of the heterochromatin, satellite II (1.680 g/cm3) hybridizes with two bands at the margin of the heterochromatin. In situ hybridization with polytene chromosomes of Chironomus thummi revealed the presence of G. barbipes satellite sequences also in the Ch. thummi genome at varios locations, mainly the centromere regions.