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Nucleic Acids Research, 1988, Vol. 16, No. 8 3437-3454
© 1988

Articles

Proteins associated with rabbit reticulocyte mRNA caps during translation as investigated by photocrosslinking

Jay R. Greenberg1 and Virginia E. Burn2

Cell Biology Group, The Worcester Foundation for Experimental Biology 222 Maple Avenue, Shrewsbury, MA 01545, USA

1To whom correspondence should be addressed. Present address: 310 Kline Biology Tower, Department of Biology, Yale University, PO Box 6666, New Haven, CT 06511, USA

Received September 4, 1987. Revised March 2, 1988. Accepted March 2, 1988.

This laboratory previously detected by UV crosslinking number of proteins associated with cytoplasnic mRNA in mammalian cells, and the data suggested that they are involved in translation. To find oat which proteins are associated with caps we made use of retionlocyte mRNA specifically labeled in the cap with 12P together with a cell-free translation system and UV crosslinking. Approximately 8 bands corresponding to proteins crosslinked to the cap itself have been detected by polyacrylamide gel electrophoresis after UV crosslinking and digestion with RNases or tobacco pyrophosphatase. All but one were specific for methylated caps. One was similar in size and partial peptide map to a cap-binding protein, CBP I, previously identified in other laboratories, and most of the others corresponded to proteins previously known to be associated with mRNA but not known to be associated with caps. The results suggest that most mRNA-associated proteins are associated with caps or poly(A). Also, the number of cap-associated proteins nay be greater than previously suspected.

2Present address: Department of Medicine, St Vincent's Hospital, 25 Winthrop Street, Worcester, MA 01604, USA


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