TY - JOUR

T1 - DNA-Protein Interactions between Mammalian Nuclear Proteins and a GCC-Element Included in a Composite cis-Acting Element of Mouse Ribosomal Protein L32 Promoter

AU - Orlov, S. V.

AU - Kuteikin, K. B.

AU - Dizhe, E. B.

AU - Kuryshev, V. Yu

AU - Shpakovich, V. M.

AU - Perevozchikov, A. P.

PY - 1999

Y1 - 1999

N2 - DNA-protein complex formation between the sequence GC(GCC)4 (GCC-element) of mouse ribosomal protein L32 (rpL32) promoter and nuclear proteins of mouse and human cells has been studied using gel retardation and South-Western blotting methods. The rpL32 promoter fragment (-24...+11) was able to form specific complexes with mouse and human nuclear proteins mainly due to the presence of the GCC- element (-19...-6). DNA-protein complex patterns exhibited marked tissue-specificity. Three nuclear polypeptides of ∼18, 28, and 50 kD that bind to the rpL32 promoter region (-24...+11) have been detected in HeLa cells by ligand blotting. At least one of them (18 kD) interacted with the GCC-element directly. The same fragment of the promoter interacted only with one nuclear polypeptide (28-31 kD) from human fibroblasts. DNA-protein complex formation between the investigated rpL32 promoter fragment containing the GCC- element and human fibroblast nuclear proteins is Zn2+-dependent. The method of functional titration (in vivo competition in the CAT-test) revealed that the GCC-element within the rpL32 promoter functions as a positive cis-acting transcriptional element in NIH 3T3 cells. Thus, our data characterize the sequence GC(GCC)4 as a functionally active cis-element included as a component in the more complex (composite) cis-element of mouse rpL32 promoter exhibiting tissue-specific properties. In various mammalian cell types the GCC-element can interact with various nuclear proteins, and the mode of these interactions can be determined by its relative position to other cis-elements in the regulatory sites of the genome.

AB - DNA-protein complex formation between the sequence GC(GCC)4 (GCC-element) of mouse ribosomal protein L32 (rpL32) promoter and nuclear proteins of mouse and human cells has been studied using gel retardation and South-Western blotting methods. The rpL32 promoter fragment (-24...+11) was able to form specific complexes with mouse and human nuclear proteins mainly due to the presence of the GCC- element (-19...-6). DNA-protein complex patterns exhibited marked tissue-specificity. Three nuclear polypeptides of ∼18, 28, and 50 kD that bind to the rpL32 promoter region (-24...+11) have been detected in HeLa cells by ligand blotting. At least one of them (18 kD) interacted with the GCC-element directly. The same fragment of the promoter interacted only with one nuclear polypeptide (28-31 kD) from human fibroblasts. DNA-protein complex formation between the investigated rpL32 promoter fragment containing the GCC- element and human fibroblast nuclear proteins is Zn2+-dependent. The method of functional titration (in vivo competition in the CAT-test) revealed that the GCC-element within the rpL32 promoter functions as a positive cis-acting transcriptional element in NIH 3T3 cells. Thus, our data characterize the sequence GC(GCC)4 as a functionally active cis-element included as a component in the more complex (composite) cis-element of mouse rpL32 promoter exhibiting tissue-specific properties. In various mammalian cell types the GCC-element can interact with various nuclear proteins, and the mode of these interactions can be determined by its relative position to other cis-elements in the regulatory sites of the genome.

KW - Cis-elements

KW - DNA-binding proteins

KW - GCC-element

KW - Mouse rpL32 gene

KW - Transcriptional factors

UR - http://www.scopus.com/inward/record.url?scp=0033070017&partnerID=8YFLogxK

M3 - Article

C2 - 10187914

AN - SCOPUS:0033070017

VL - 64

SP - 207

EP - 212

JO - Biochemistry (Moscow)

JF - Biochemistry (Moscow)

SN - 0006-2979

IS - 2

ER -