Transcription of the alpha A-globin gene of the duck. Development of a homologous in vitro system and identification of trans-acting factors.
A homologous in vitro transcription system was developed in which the alpha A-globin gene of the duck was faithfully transcribed. Whole-cell extracts from duck erythrocytes were separated into fractions A, B, C and D by consecutive elution from phosphocellulose columns and were individually reconstituted in run-off transcription assays. Fractions A, C and D were required to achieve faithful initiation on the alpha A-globin gene. The latter fractions were mutually interchangeable with comparable fractions from HeLa cells. A fourth fraction, B, was not required but enhanced basal transcription when reconstituted with fractions A, C and D or a very low amount of HeLa whole-cell extract which by itself did not yield a detectable signal. Fraction B from duck erythrocytes was further purified by chromatography on DEAE-Sephadex and was shown to contain two trans-acting factors. One of these differentially acts on the alpha A-globin gene of the duck. The other component from duck erythrocytes surprisingly resembles the upstream stimulatory factor, previously isolated from HeLa cells. This latter protein binds to and trans-activates the adenovirus 2 major late promoter, but is not involved in the transcription of the alpha A-globin gene.
Weingart
S
S
Institut für Molekularbiologie und Tumorforschung der Phillipps-Universität Marburg, Federal Republic of Germany.
Sommer
U
U
Gerhold
H
H
Seifart
K H
KH
D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D003850:Deoxyribonuclease I; D003851:Deoxyribonucleases; D004372:Ducks / Q000235:genetics*; D004912:Erythrocytes / Q000378:metabolism; D005786:Gene Expression Regulation; D015696:Gene Products, tat; D005914:Globins / Q000096:biosynthesis / Q000235:genetics*; D006367:HeLa Cells / Q000378:metabolism; D006358:Hot Temperature; D006868:Hydrolysis; D010957:Plasmids; D011485:Protein Binding; D014157:Transcription Factors / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic
D015696:Gene Products, tat; D014157:Transcription Factors; D005914:Globins; D003851:Deoxyribonucleases; D003850:Deoxyribonuclease I