Differential RNA expression and polyribosome loading of alternative transcripts of the Akap4 gene in murine spermatids.
An X chromosome-linked gene, Akap4, is expressed only during spermiogenesis and encodes the major fibrous sheath protein of the mouse sperm flagellum. All sperm contain the AKAP4 protein even though only X chromosome-bearing spermatids express the gene, indicating that the Akap4 mRNA and/or protein must be shared among the conjoined spermatids via the intercellular bridges. There are two mouse Akap4 cDNA clones, Akap82 and Fsc1, which represent mRNAs that arise by alternative processing of a single gene. Although Akap82 and Fsc1 encode identical mature proteins, they differ in their 5' UTRs. We hypothesized that the expression pattern of these two mRNAs might be relevant to the issue of mRNA and/or protein transport into adjacent spermatids. Expression of both transcripts began in round spermatids, but the amount of the Akap82 transcript in condensing spermatids increased twofold relative to Fsc1. Significantly, only the Akap82 transcript was found on polyribosomes and translated in spermatids. These results indicate that the Akap82 transcript and/or its protein must be shared among the conjoined X and Y chromosome-bearing spermatids. Although Fsc1 was not polysomal, both the Akap82 and Fsc1 transcripts were deadenylated during spermiogenesis, suggesting that deadenylation is not always correlated with loading of mRNAs onto polyribosomes in germ cells. The distinct 5' UTR sequences in Akap82 and Fsc1 did not differ in their ability to regulate translation of reporter constructs either in vivo or in vitro. Antisense RNA transcripts complementary to both the Akap82 and Fsc1 mRNAs were present, suggesting that translatability may be regulated by these RNAs.
Nipper
Rick W
RW
Center for Research on Reproduction and Women's Health, University of Pennsylvania Medical School, Philadelphia, Pennsylvania 19104, USA.
Chennothukuzhi
Vargheese
V
Tutuncu
Levent
L
Williams
Carmen J
CJ
Gerton
George L
GL
Moss
Stuart B
SB
D020121:5' Untranslated Regions; D054758:A Kinase Anchor Proteins; D017398:Alternative Splicing; D000818:Animals; D008297:Male; D051379:Mice; D011132:Polyribosomes / Q000378:metabolism*; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics*; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D032961:Sperm Midpiece / Q000378:metabolism; D013087:Spermatids / Q000378:metabolism*; D014960:X Chromosome / Q000378:metabolism; D014998:Y Chromosome / Q000378:metabolism
D020121:5' Untranslated Regions; D054758:A Kinase Anchor Proteins; C485479:Akap4 protein, mouse; D011498:Protein Precursors; D016372:RNA, Antisense; D012333:RNA, Messenger