Document Detail

Red cell ghost-mediated microinjection of RNA into HeLa cells. II. Cellular translation of protamine mRNA; post-translational modifications and nuclear binding of newly-synthesized protamine.
MedLine Citation:
PMID:  6825718     Owner:  NLM     Status:  MEDLINE    
Red cell ghosts loaded with protamine messenger RNA (pmRNA) were fused to HeLa cells using polyethylene glycol, as a means of introducing the mRNA into heterologous cells. The recipient cells were capable of translating the RNA into the three protamine polypeptides, which may be resolved as three peaks (CI, CII, and CIII) by cation exchange chromatography. The synthesis of components CII and CIII was easily observed with possible traces of CI as well. The HeLa cells also phosphorylated CII after synthesis. However, this phosphorylation did not occur with CIII. In addition, CII but not CIII localized in the nucleus of the HeLa cells after synthesis. Thus, a correlation of post-translational modification with nuclear entry was observed. Localization in the nucleus, however, was not accompanied by the same tight binding of protamine to chromatin as is seen in the homologous trout testis spermatid cells. In the spermatid cells, protamine elutes from chromatin at a salt concentration of 1.2 M NaCl. In contrast, in the HeLa cells, the newly synthesized CII which had entered the nucleus, could be eluted with 0.6 M NaCl. Thus, the tight binding of protamine to chromatin in trout testis may require a series of concomitant developmental events, such as core histone hyper-acetylation (Christensen, M E & Dixon, G-H. In press) [17], which would be lacking in the HeLa cells.
C Boogaard; G H Dixon
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Experimental cell research     Volume:  143     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1983 Jan 
Date Detail:
Created Date:  1983-04-07     Completed Date:  1983-04-07     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  191-205     Citation Subset:  IM    
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MeSH Terms
Cell Compartmentation
Erythrocyte Membrane*
Hela Cells
Microinjections / methods*
Protamines / biosynthesis*
Protein Biosynthesis
Protein Processing, Post-Translational*
RNA, Messenger / administration & dosage*,  metabolism
Time Factors
Reg. No./Substance:
0/Protamines; 0/RNA, Messenger

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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