Immobilized polynucleotides and nucleic acids.
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The effect of the ligts-7 mutation on cell survival and the extent of DNA repair after UV (254 nm) irradiation was determined for wild-type and uvrB5 cells of E. coli K-12 at 30 degrees and 42 degrees C. At the restrictive temperature (42 degrees C) the ligts-7 mutation resulted in (i) a decrease in the extent of repair of DNA incision breaks arising during the excision repair process, and (ii) a decrease in the extent of post-replicational repair of gaps in newly-synthesized DNA. These deficiencies in DNA repair correlated with increases in cellular sensitivity to killing by UV radiation. Thus, DNA lagase plays an important role in vivo in both the excision and post-replicational repair processes.
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The binding of adenine derivatives to Na(+)-montmorillonite increases in the order 5'AMP, 3'-AMP, 5'ADP < adenosine < purine, adenine. With the exception of cytosine, cytosine derivatives bind less strongly than the corresponding adenine derivatives in the order 5'-CMP < cytidine < cytosine. There is little difference in the binding of uracil derivatives and these compounds bind less strongly than the corresponding adenine analogs. It is concluded that the adenine ring in adenine derivatives is protonated by the acidic montmorillonite surface and binding is a consequence of the electrostatic interaction between the protonated base and the negative charges on the surface of the montmorillonite. Different binding trends were observed with Cu(2+)-montmorillonite with AMP binding more strongly than adenosine and UMP binding more strongly than uridine. It is concluded that ligation to the Cu2+ is a major force in the binding of nucleotides to Cu(2+)-montmorillonite and are not readily washed from the clay. Factors contributing to the binding are discussed. Watson-Crick hydrogen bonding of 5'-AMP to poly(U) and 5'GMP to poly(C) was observed when the homopolymers are bound to the surface of the clay. No association of 5'-UMP to poly(U) bound to clay was detected. The possible role of montmorillonite clays in the prebiotic formation of RNA is discussed.
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Nine human-human hybridoma clones, secreting monoclonal antibodies reactive with nucleic acids, were generated by fusing with lymphocytes of lung cancer or systemic lupus erythematosus patients. These hybridoma antibodies were classified into 5 types, in terms of reactivities with DNA, RNA, various synthetic nucleic acids and cardiolipin. Hybridoma clone SU-1 secreted antibody reacting with dsDNA, ssDNA and RNA (type I). Clone HL-321 did not react with these, but with poly (dT), poly (I) and poly (G) (type II). Clone HL-349 was reactive with almost all nucleic acids tested and also with cardiolipin (type III). Clones HF-4, HF-7, HB-7 and HL-259 reacted with ssDNA, poly (A), poly (G) and cardiolipin, but not with RNA (type IV). HB-5 and SH-9 antibodies were reactive only with poly (dT) (type V).
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