Silkworm TFIIIB binds both constitutive and silk gland-specific tRNA Ala promoters but protects only the constitutive promoter from DNase I cleavage.
We have identified a complex between TFIIIB and the upstream promoter of silkworm tRNA Ala genes that is detectable by gel retardation and DNase I footprinting. Formation of this complex depends on the integrity of previously identified upstream promoter elements and on the presence of other silkworm transcription factors, either TFIIID or a fraction that contains both TFIIIC and TFIIID. We have used this complex to compare the interactions of TFIIIB with two kinds of tRNA Ala genes whose different in vitro transcription properties are conferred by the upstream segments of their promoters. These are the tRNA C Ala genes, which are transcribed constitutively, and the tRNA SG Ala genes, which are transcribed only in the silk gland. We find that TFIIIB binds tRNA SG Ala genes with lower affinity than it binds tRNA C Ala genes. In addition, the TFIIIB complex formed on tRNA SG Ala genes differ qualitatively from those formed on tRNA C Ala genes. Both the transcriptional activity of tRNA SG Ala complexes and the ability of the complexes to protect upstream DNA from DNase I digestion are reduced.
Young
L S
LS
Institute of Molecular Biology, University of Oregon, Eugene 97403-1229, USA.
Ahnert
N
N
Sprague
K U
KU
D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000378:metabolism*; D003001:Cloning, Molecular; D003850:Deoxyribonuclease I / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012344:RNA, Transfer, Ala / Q000235:genetics* / Q000378:metabolism; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors / Q000378:metabolism*
D012344:RNA, Transfer, Ala; D035361:Transcription Factor TFIIIB; D014157:Transcription Factors; D003850:Deoxyribonuclease I