Greetings, thank you for your patience. We are aware that the northern data
does not coincide with the complementation analysis. However, we are
currently repeating the northern analysis to determine if the 'extra'
altered transcript in 2A is just an artifact. If you look at the northern,
83A appears to have a stronger signal at 4kb when compared to 2A. The
complementation study does indeed point out that the mutation in 2A is in a
different gene from that of 83A. We are confident that the mutation is in
the 83A mutant. Thus, we are changing the following statement in the
abstract (<up></up> = superscript):
'In addition, a 4 kb transcript was seen in both mutant lines suggesting an
alteration.'
to
'In addition, a 4 kb transcript was seen in the AT7683A mutant, which
suggests an alteration.'
We would like to use the symbol Ubc87F, the name Ubiquitin conjugating
enzyme, and the map location 87F12 for this gene.
I hope this clears any confusion,
Alexander Runk
Subject: RE: EDRC99 - abstract ID140 addendum
Ok, here's an up-to-date title and abstract that we will be using in the
conference (<i> = indentation).
Analysis of a Novel Muscle Ubiquitin Conjugating Enzyme in <i>Drosophila</i>
Our goal is to characterize mutations in novel <i>Drosophila</i> genes that
disrupt the structure and function of the neuromuscular system in a manner
analogous to the known myopathies in humans. A homozygous enhancer-trap
line, AT76, was generated resulting in a stable insertion of P{lwB} in the
genome. Cytogenetic and P1 mapping placed the insertion in the region 87F12.
These flies display wild-type flight behavior, which is consistent with the
normal indirect flight muscle (IFM) morphology observed in electron
micrographs. Excision of the enhancer trap resulted in two recessive lethal
lines, AT7683A and AT762A, which fall within two complementation groups.
Furthermore, AT7683A displays dominant flight-defective behavior. The IFM
ultrastructure of the flight-defective flies is markedly disrupted. We
isolated a 5.5 kb genomic fragment flanking the insertion, initiated a
chromosomal walk, and cloned a 531 bp partial cDNA with a predicted amino
acid sequence that has 75% identity to the human skeletal muscle ubiquitin
conjugating enzyme, ube2g, an E2 class enzyme. RFLP analysis shows
chromosomal rearrangement within the region of the cDNA. Northern blots
probed with this cDNA reveal 850 bp, 1.3 kb, and 2 kb transcripts. In
addition, a 4 kb transcript was seen in the AT7683A mutant, which suggests
an alteration. A 9 kb genomic fragment containing this putative gene was
cloned into pP{CaSpeR} to perform germline transformations. Currently, we
are analyzing the transformants to determine whether the mutant phenotypes
have been rescued.
Sincerily,
Alexander Runk
\-----Original Message-----
Sent: Thursday, September 23, 1999  1:57  AM
Subject: Re: EDRC99 - abstract ID140
Dear Runk,
Just over a week ago, I sent you a query concerning your abstract for
the upcoming EDRC99 conference. I was wondering if you'd had a chance
to look at it. I enclose the mail below for your convenience.
Thanks
Chihiro
>
> Dear Dr. Runk,
>
> We are currently curating the abstracts for the upcoming European
> Drosophila Research Conference in Zurich, for FlyBase. I am writing in
> connection with your abstract:
>
> 'Analysis of a Novel Muscle-Specific Ubiquitin-Conjugating Enzyme in
> Drosophila'
>
> In you abstract you talk about two lines derived from the enhancer trap
line AT76, which you call 'AT7683A' and 'AT7683A' (<up></up> = superscript).
>
> You state that these 'fall within two complementation groups.' which
> suggests that they are in two separate genes. However, later in you
> abstract you state 'a 4 kb transcript was seen in both mutant lines
> suggesting an alteration in the mRNA.' which suggests that the two
> lines are mutants in the same gene.
>
> Could you confirm whether these two mutant lines are mutations within
> the same gene or two different ones.
>
> Also, for 'AT7683A' you state that the mutation is in a homologue of
> 'human ubiquitin-conjugating enzyme (ube2g)'. Would you like the
> symbol 'Ube2g' to be the valid symbol for this gene in FlyBase, or do
> you have an alternative symbol for the gene that is mutated in
> 'AT7683A' ?
>
> Thank you for your help,
>
> with best wishes,
>
> Chihiro
> ----------------------------------------------------------------------
> Chihiro Yamada.
>
> FlyBase (Cambridge),
> ----------------------------------------------------------------------
>