Characterization of the expression and regulation of MK5 in the murine ventricular myocardium.
MK5, a member of the MAPK-activated protein kinase family, is highly expressed in the heart. Whereas MK2 and MK3 are activated by p38 MAPK, MK5 has also been shown to be activated by ERK3 and ERK4. We studied the regulation of MK5 in mouse heart. mRNA for 5 splice variants (MK5.1-5.5), including the original form (MK5.1), was detected. MK5 comprises 14 exons: exon 12 splicing was modified in MK5.2, MK5.3, and MK5.5. MK5.2 and MK5.5 lacked 6 bases at the 3'-end of exon 12, whereas MK5.3 lacked exon 12, resulting in a frame shift and premature termination of translation at codon 3 of exon 13. MK5.4 and MK5.5 lacked exons 2-6, encoding kinase subdomains I-VI, and were kinase-dead. All 5 MK5 variants were detected at the mRNA level in all mouse tissues examined; however, their relative abundance was tissue-specific. Furthermore, the relative abundance of variant mRNA was altered both during hypertrophy and postnatal cardiac development, suggesting that the generation or the stability of MK5 variant mRNAs is subject to regulation. When expressed in HEK293 cells, MK5.1, MK5.2 and MK5.3 were nuclear whereas MK5.4 and MK5.5 were cytoplasmic. A p38 MAPK activator, anisomycin, induced the redistribution of each variant. In contrast, MK5 co-immunoprecipitated ERK3, but not ERK4 or p38 alpha, in control and hypertrophying hearts. GST pull-down assays revealed unbound ERK4 and p38 alpha but no free MK5 or ERK3 in heart lysates. Hence, 1) in heart MK5 complexes with ERK3 and 2) MK5 splice variants may mediate distinct effects thus increasing the functional diversity of ERK3-MK5 signaling.
Dingar
Dharmendra
D
Montreal Heart Institute, 5000 Belanger St., Montréal, Québec, Canada.
Benoit
Marie-Josée
MJ
Mamarbachi
Aida M
AM
Villeneuve
Louis R
LR
Gillis
Marc-Antoine
MA
Grandy
Scott
S
Gaestel
Matthias
M
Fiset
Celine
C
Allen
Bruce G
BG
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D006332:Cardiomegaly / Q000201:enzymology / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D006321:Heart / Q000254:growth & development; D006352:Heart Ventricles / Q000201:enzymology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D020935:MAP Kinase Signaling System; D051379:Mice; D008810:Mice, Inbred C57BL; D048053:Mitogen-Activated Protein Kinase 6 / Q000378:metabolism; D008969:Molecular Sequence Data; D009206:Myocardium / Q000201:enzymology*; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism
D047908:Intracellular Signaling Peptides and Proteins; D012333:RNA, Messenger; C111049:MAP-kinase-activated kinase 5; D017346:Protein Serine-Threonine Kinases; D048053:Mitogen-Activated Protein Kinase 6