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PMID: 6206497 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Expression of specific mRNAs during adipose differentiation: identification of an mRNA encoding a homologue of myelin P2 protein.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 17 ·1984-09-00 ·Pages 5468-72

Bernlohr DA, Angus CW, Lane MD, Bolanowski MA, Kelly TJ

Abstract

To identify and characterize specific mRNAs that increase in abundance during differentiation of mouse 3T3-L1 preadipocytes, a cDNA library was constructed from poly(A)+RNA isolated from differentiated 3T3-L1 adipocytes. Mixed probe isotope ratio selection and RNA blot analyses have identified several unique cDNA clones that represent mRNA species expressed either exclusively or at dramatically increased levels in differentiated cells. Further characterization of one such clone (pAL422) revealed that the corresponding mRNA, detectable only after differentiation, is approximately the same length (600 +/- 150 bases) as the cDNA insert (672 bases). The complete nucleotide sequence of the cDNA insert in pAL422 revealed a single long open reading frame that encodes a 132 amino acid polypeptide (the 422 protein) of 14.6 kDa. These and other results suggest that this cDNA may represent a nearly full-length copy of the mRNA. Computer-assisted analyses showed that the 422 protein shares 69% and 64% homology with myelin P2 proteins from rabbit and bovine peripheral nerves, respectively, as well as 23% and 30% homology with fatty-acid binding proteins from rat liver and intestine, respectively. Moreover, the mRNA hybrid selected by pAL422 DNA directs the in vitro translation of an approximately equal to 13 kDa polypeptide, and this protein is specifically immunoprecipitated by antiserum against bovine myelin P2. These observations strongly suggest that the 422 protein is a structural, and possibly functional, analog of myelin P2.

MeSH Terms
Adipose Tissue/cytology,metabolism Amino Acid Sequence Animals Base Sequence Cell Differentiation Cell Line Cloning, Molecular DNA/analysis Mice Myelin Basic Protein/genetics Myelin P2 Protein Nucleic Acid Hybridization Protein Biosynthesis RNA, Messenger/genetics Templates, Genetic
Chemicals
Myelin Basic Protein Myelin P2 Protein RNA, Messenger DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bernlohr D A
Angus C W
Lane M D
Bolanowski M A
Kelly T J
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40 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.81.17.5468
Published
1984-09-00
Pages
5468-72
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC391726
Subset
IM
Grants
NIADDK NIH HHS · 1 F32 AM06954 · United States
NCI NIH HHS · 5 PO1 CA16519 · United States
NIADDK NIH HHS · AM14574 · United States
Databases
GENBANK
K02109
External Links
PubMed source DOI: 10.1073/pnas.81.17.5468 PMC Full Text View journal details
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