Sequence-specific DNA-binding proteins are components of a nuclear matrix-attachment site.
We have identified a nuclear matrix-attachment region within an upstream element of a human H4 histone gene promoter. Nuclear matrix proteins, isolated and solubilized from HeLa S3 cells, were found to interact with sequence specificity at this matrix-attachment region. Several types of assays for protein-DNA interaction showed that the minimal sequence for the nuclear matrix protein-DNA interaction was 5'-TGACGTCCATG-3'; the underlined region corresponds to the core consensus sequence for ATF transcription factor binding. Two proteins with molecular masses of 43 and 54 kDa were identified by UV-crosslinking analysis as integral components of this protein-DNA complex. The molecular masses of these proteins and the ATF-binding site consensus sequence suggest that these proteins are members of the ATF family. Our results provide direct evidence for nuclear matrix localization of sequence-specific DNA-binding factors for an actively transcribed gene. The proximity of a strong positive transcriptional regulatory element to the matrix-attachment region of this gene suggests that the nuclear matrix may serve to localize and concentrate trans-acting factors that facilitate regulation of gene expression.
Dworetzky
S I
SI
Department of Cell Biology, University of Massachusetts Medical Center, Worcester 01655.
Wright
K L
KL
Fey
E G
EG
Penman
S
S
Lian
J B
JB
Stein
J L
JL
Stein
G S
GS
D001483:Base Sequence; D001665:Binding Sites; D016384:Consensus Sequence; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015530:Nuclear Matrix / Q000648:ultrastructure*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping
D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D006657:Histones; D009687:Nuclear Proteins