THE INTERACTION BETWEEN EXOGENOUS DNA AND SPERM CELLS

被引:99
|
作者
LAVITRANO, M
FRENCH, D
ZANI, M
FRATI, L
SPADAFORA, C
机构
[1] CNR,INST BIOMED TECHNOL,VIA GB MORGAGNI 30-E,I-00161 ROME,ITALY
[2] UNIV ROME LA SAPIENZA,DEPT EXPTL MED,I-00185 ROME,ITALY
关键词
SPERM CELL; RECOMBINANT DNA; FERTILIZATION; GENETIC TRANSFORMATION; TRANSGENIC ANIMALS;
D O I
10.1002/mrd.1080310302
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Epididymal sperm cells, incubated with plasmid DNA, showed a spontaneous tendency to interact with the exogenous nucleic acid. We have investigated the molecular basis of such interaction. Exogenous DNA is taken up by sperm cells over a 15- to 20-min period and is specifically localized on the nuclear area of the sperm head. DNA was reversibly bound to spermatozoa since it can be competed out by excess of cold competitor DNA or by other polyanions as heparin and dextran sulphate. By contrast, poly-L-lysine, a polycation, favours the uptake. DNA molecules of large size (7 kb) were preferentially taken up as compared to smaller ones (150-750 bp). Acidic proteins were also taken up and concentrated, as for DNA, at the nuclear level. These data strongly suggested that ionic interactions may occur between foreign molecules and a substrate located in the sperm head. On the basis of Southwestern analysis, a sperm head protein(s) ot 30-35 KD is identified as potential substrate for exogenous DNA binding. Moreover, we have found that seminal plasma contains factor(s) which abolish sperm permeability, exerting a powerful inhibitor effect on DNA uptake. The presence of a specific binding protein for the DNA and of a factor inhibiting such interaction support the existence of a mechanism controlling, through specific factors, the sperm - DNA interaction.
引用
收藏
页码:161 / 169
页数:9
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